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Page 1

Sample Preparation

A Guide to Methods and Applications

A SPG Table of Contents

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Page iTrademarks ....................................................... vi

One

Preliminary
Te c h n i q u e s

Introduction ...................................................... vii

Particulate Removal .......................................... 1
Buffer and Reagent Ultrapurification ............... 3
Ionic Contaminants ..............................................4
UV Absorbing and Polar Compounds....................4
Endotoxins ...........................................................4
References ............................................................5

Two

Desalting andBuffer Exchange

Protein Concentration and Enrichment ........... 6
Dilute Proteins ......................................................7
Acidic and Basic Proteins ......................................7
Serum Proteins .....................................................9Albumin............................................................9
Antibodies ........................................................9
References ..........................................................13

Introduction ..................................................... 17
Methods ........................................................... 18
MW > 2,000 Daltons (Gel Filtration) ...................18
MW < 2,000 Daltons (Ion Exchange andAdsorption) ........................................................18

Three

Plasmid
Purification andProbe Clean-up

References........................................................ 20

Introduction ..................................................... 21
Applications ..................................................... 22
Ethidium Bromide and Propidium
Iodide Removal...................................................22
DNA Isolation from Agarose Gel .........................22
Radioactive Label Removal..................................22
Plasmid Purification ............................................23
Removal of PCR Inhibiting Contaminants............25
References........................................................ 25i

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Page ii

Detergent
RemovalIntroduction ..................................................... 27
Methods ........................................................... 28
Non-Ionic Detergents .........................................28
Ionic Detergents .................................................28
Sensitive or Dilute Protein Samples .....................28

Five

Ionic
ContaminantRemovalReferences........................................................ 29

Introduction ..................................................... 31
Deionization..................................................... 31
Mechanism.........................................................32
Methods.............................................................32
Applications........................................................33
Reagents, Buffers, and Water...........................33
Protein Samples ..............................................33
Carbohydrate Samples ....................................33
References ..........................................................33
Metal Ion Removal........................................... 34
Mechanism.........................................................35
Metal Cations .................................................35
Metal Anion Complexes ..................................35
Metal Cation and Anion Removal ....................35
Applications........................................................35
Trace Metal Removal, Concentration,and Metal Analysis ..........................................35
Buffer Purification ...........................................36
References ..........................................................36
Anion and Cation Removal .............................. 38
Resin Selection Parameters .................................39
pH ..................................................................40
Ionic Form ......................................................40
Porosity ..........................................................40
Particle Size.....................................................40
Time and Temperature ...................................40(continued on next page)ii

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Applications........................................................41
Ampholyte Removal........................................41
Organic Acids .................................................42
Peptides and Amino Acids...............................42
Sugars and Sugar Alcohols ..............................42
References ..........................................................42

Six

Organic
CompoundRemoval andConcentration

Introduction ..................................................... 49
Mechanisms ..................................................... 50
Hydrophobic Interaction.....................................50
Gel Permeation...................................................50
Ion Exchange......................................................50
Applications ..................................................... 50
Biological Applications........................................50
Environmental Applications ................................51
Pesticides, PCB, and PAH ................................51

Seven

Columns andAccessories

References........................................................ 51

Introduction ..................................................... 55
Columns ........................................................... 55
Small Volume .....................................................55
Bio-Spin Columns ...........................................55
Poly-Prep Columns..........................................56
Intermediate Volume ..........................................56
Econo-Pac Columns ........................................56
Large Volume .....................................................57
Econo-Column Columns .................................57
Systems and Accessories .................................. 57
Econo System .....................................................57
Flow Adaptors ....................................................58
Tubing and Fittings ............................................58
Poly Column Rack...............................................58

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Page iv

Product
Information

Membrane Filters for Particulate Removal...... 59
Prep-Disc Membrane Filters ................................59
Micro Prep-Disc Membrane Filters ......................59
Ion Exchange Chromatography Resins ........... 59
Analytical Grade Resins .......................................59
Prefilled Poly-Prep Columns ................................60
Biotechnology Grade Resins................................61
Molecular Biology Grade Resins ..........................61
Macro-Prep Ion Exchange Supports ....................61
Econo-Pac Cartridges..........................................62
Bio-Gel A Ion Exchangers....................................62
Hydrophobic Interaction Chromatography
Supports........................................................... 62
Macro-Prep Supports..........................................62
Econo-Pac Cartridges..........................................63
Bio-Beads SM Adsorbents ...................................63
Gel Filtration Chromatography Supports........ 63
Bio-Gel P Polyacrylamide Gels.............................63
Econo-Pac Columns and Cartridges ....................64
Bio-Gel A Agarose Gels .......................................64
Bio-Beads S-X Beads ...........................................64
Affinity Chromatography Supports ................. 64
Ready-to-Use Affinity Supports............................64
Activated Affinity Supports..................................65
Carbodiimide Activated Supports........................65
Econo-Pac Columns and Cartridges ....................66
Hydroxyapatite Chromatography Supports.... 66
Bio-Gel Hydroxyapatite Media ............................66
Macro-Prep Hydroxyapatite Supports .................66
Econo-Pac Cartridges..........................................66
DNA Purification Supports .............................. 67
Prep-A-Gene DNA Purification Reagents .............67
InstaGene DNA Purification Matrix .....................67
Chromatography Columns .............................. 67
Standard Econo-Column Columns ......................67
Standard Jacketed Econo-Column Columns ........69(continued on next page)iv

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Open-Ended Jacketed Econo-Column Columns...69
Flow Adaptors ....................................................69
Glass Reservoirs ..................................................69
Econo-Column Funnels.......................................69
Bio-Spin Columns ...............................................69
Poly-Prep Columns .............................................69
Econo-Pac Columns............................................70

Nine

Literature
Available

Econo Systems and Accessories ....................... 70
Econo Systems....................................................70
Tubing ...............................................................70
Poly Column Rack...............................................70

Literature Available.......................................... 71v

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Page vi

Tr a d e m a r k s
The following trademarks and registered trademarks used in this guide are theproperty of Bio-Rad Laboratories.
Affi-Gel®
Affi-Prep®
AG®
Bio-Beads®
Bio-Gel®
Bio-Rad®
Bio-Rex®

Bio-Sil®
Bio-Spin®
Chelex®
Econo-Column®
Econo-Pac®
GS Gene Prep®
InstaGene™

Macro-Prep®
MAPS®
Poly-Prep®
Prep-A-Gene®
Prep-Disc®

The following trademarks are the property of the companies listed.
PCR is covered by U. S. patent number 4,683,202, issued to Cetus Corporation
Cibacron is a trademark of Ciba-Geigy
Dowex is a trademark of Dow Chemical Company
Norprene, Fluran, PharMed and Tygon are trademarks of Norton Co
Amberlite, Plexiglas, Triton, Tween, and XAD are trademarks of Rohm and Haas
Companyvi

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Page vii

Introduction
Sample preparation involves a wide range of techniques which, in most cases, makeuse of small chromatographic columns, filters, and media to transform a sample
which cannot be directly analyzed into one that fits the requirements of the analytical technique to be used. In the past, sample preparation techniques have been mostclosely associated with classical chromatographic analysis. Currently there is a growing interest in sample preparation for other analytical methods in molecular biology
and biotechnology. This guide will assist the biochemist or molecular biologist in
selecting appropriate sample preparation methods. It covers common techniquesin sample preparation and explains separation strategies and alternatives.
Bio-Rad offers a complete line of products for sample preparation applications, anda technical staff to assist you in designing an effective protocol for your application.
For more information on products or applications, contact your Bio-Rad representative, or, in the US, call 1-800-4BIORAD and press 2 for technical service.

vii

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Page viii

viii

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Page 1Preliminary Techniques
Particulate Removal
Sample with particulates

Prefilter

Clarification

Filtration

Sterilization

Volume > 2 ml
Prep-Disc filter
5 µm

Volume > 2 ml
Prep-Disc filter
1 µm

Volume > 2 ml
Prep-Disc filter
0.45 µm

Volume > 2 ml
Prep-Disc filter
0.2 µmVolume < 2 ml
Micro
Prep-Disc filter
5 µm

Volume < 2 ml
Micro
Prep-Disc filter
1 µm

Volume < 2 ml
Micro
Prep-Disc filter
0.45 µm

Volume < 2 ml
Micro
Prep-Disc filter
0.2 µmTable 1.1. Guide to Particulate Removal
Filtration for particulate removal is the simplest and most common sample preparation procedure. It is commonly used before sample injection to minimize
particulate clogging of lines and valves, to protect valuable HPLC columns, and tomaintain the integrity of liquid chromatography columns. Filtration is also used
for the clarification of liquids, sterilization of tissue culture media and other liquids, and ultracleaning of water and alcohols.
Prefiltration, clarification, filtration, and sterilization techniques are defined by thesize of the particles removed from solution (Table 1.1). Prefiltration is the removalof large particulates (>5 µm), clarification is the removal of visible particulates (>1 µm), filtration is the removal of most particulates (> 0.45 µm), and sterilization
is filtration that also removes bacterial contaminants (> 0.2 µm). The 0.45 µm
pore size is the most commonly used filter and is employed in most HPLC applications. Prefiltration is also highly recommended to remove large particles that may
clog smaller pore size filters.
Bio-Rad offers two filter types, the Prep-Disc and Micro Prep-Disc filters. Each typeis available in four pore sizes (5, 1, 0.45, and 0.2 µm). The filters are syringe mountable and provide quick and easy particulate removal. They may be autoclaved for1

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Page 2sterile use, and offer a high surface area for fast flow rates. The filters are non-contaminating, and contain no extractables detected with UV absorbance at 190–370 nm.

Membrane filters.

Table 1.2. Membrane Filter Specifications
Prep-Disc Filter

Micro Prep-Disc Filter

Construction

Polypropylene housing;
PTFE membrane

Polypropylene housing;
PTFE membrane

Dimensions

2.5 cm long x 2.5 cm diameter

2.0 cm long x 0.3 cm diameter

Connections
Inlet
OutletFemale luer lock with locking tabs
Male luer taper

Female luer lock with locking tabs
Male luer taper

Retention volume(with air purge)< 0.5 ml

< 10 µl

Process volume

2–100 ml

< 2 mlSolvent and sample
compatibility

Organic solutions, acids, and bases

Organic solutions, acids, and bases

Filter area

4.3 cm2

0.07 cm2

2

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Page 3

Buffer and Reagent Ultrapurification
UV absorbing
organic
contaminants

Bio-Beads SM-2
adsorbents

Endotoxins

Affi-Prep
polymyxin gel

Contaminated bufferor reagent

Ionic
contaminant
in non-ionic
reagent

AG 501-X8 resin

Anionic
contaminant

AG 1-X8 resin

Cationic
contaminant

AG 50W-X8 resin

Metal cation
contaminantChelex 100 resin

Ionic contaminants

Polar compounds
in organic solvents

Bio-Beads SM
adsorbents

Table 1.3. Guide to Buffer and Reagent Ultrapurification
In the purification and separation of proteins, extremely pure buffers must be used
to obtain optimal results. High buffer purity is required for electrophoresis, HPLC,and sensitive enzyme studies. For example, in electrophoresis applications, urea
and formamide frequently must be deionized prior to use. Deionization removes
impurities that can cause band broadening, high background noise, and gel artifacts. Similarly, buffers contaminated with heavy metal ions and UV absorbing
impurities can cause high background noise and unstable baselines with the sensitive detectors used in HPLC analysis.

3

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Page 4Ionic Contaminants
Ionic contaminants in buffers and reagents are easily removed using ion exchange
chromatography. Strong anion and cation exchange resins, such as AG 1-X8 resin
and AG 50W-X8 resin, are used for reagent clean-up, while Chelex 100 chelating
resin is used to purify aqueous solutions and buffers.
AG 501-X8 mixed bed resin contains equivalent amounts of AG 1-X8 and AG
50W-X8 resins, making this exchanger ideal for general purpose deionization of
non-ionic reagents such as urea, glyoxal, formamide, and acrylamide. Deionization
is fast and easy; contaminated reagents can be completely deionized within anhour using simple batch techniques.
Buffer purification is easily accomplished using Chelex 100 chelating resin. Chelex
100 resin is highly selective for polyvalent cations, making it uniquely suited to
specifically remove heavy metals from aqueous solutions and buffers without altering the concentration of monovalent ions. Chelex resin has been used to purify
many buffers, including HEPES-KCl, Tris-HCl, PIPES, and phosphate buffers.

UV Absorbing and Polar Compounds
Neutral macroporous polymeric beads, like Bio-Beads SM-2 adsorbents, are usefulfor removing UV absorbing impurities from buffers and unbound fluorescent dyes
from labeled antibodies. Bio-Beads SM-2 and SM-4 non-polar polystyrene
adsorbents are particularly useful for the removal of non-polar substances or surface-active agents from aqueous solutions. Bio-Beads SM-7 adsorbents are slightly
more polar than the other types of Bio-Beads adsorbents and can adsorb polar
compounds from non-polar solvents or non-polar compounds from polar solvents.

Endotoxins
Endotoxins, pyrogenic lipopolysaccharides of gram negative-bacteria, are widespreadcontaminants of aqueous and physiological solutions. The removal of endotoxinsfrom solutions intended for biological applications is especially crucial for in vivo
applications. The Affi-Prep polymyxin support binds endotoxin molecules with highcapacity and selectivity. The support is pressure rated to 1,000 psi and can be sanitized and regenerated, making it ideal for both research and process applications.

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Page 5

References
Application

Product

Reference

Buffer clean-up

Chelex resin

Jefferson, J. R., et al., Biochem., 29, 6687 (1990);
Kalyanaraman, B., et al., Biochem., 28, 4839 (1989); Kupke,
D. W. and Fox, J. W., Biochem., 28, 4409 (1989); Pai, S-C.,
et al., Anal. Chem., 62, 774 (1990); Shang, Z., et al.,Biochem., 28, 9790 (1989); Shrake, A., et al., Biochem., 28,
6281 (1989).

HEPES, KCl

Chelex resin

Crouch, T. H. and Klee, C. B., Biochem., 19, 3692 (1980).

MOPS

Chelex resin

Tsai, T.-C., et al., Biochem., 24, 3180 (1985).

PIPES-KCl

Chelex resin

Swanson, J. E. and Feigenson, G. W., Biochem., 29, 8291
(1990).

Potassium phosphate

Chelex resin

Oritz de Montellano, P. R. and Grab, L. A., Biochem., 26,5310 (1987).

Sodium phosphate

Chelex resin

Kaplan, B. B., et al., Biochem., 17, 5516 (1978); Reiss, P. D.,
et al., Anal. Biochem., 140, 162 (1984).

TBS

Chelex resin

Busby, T. F. and Ingham, K. C., Biochem., 26, 5564 (1987).

TES, NaCl

Chelex resin

Burger, D., et al., Biochem., 23, 1966 (1984).

bis-Tris

Chelex resin

Gupta, R. K., J. Biol. Chem., 251, 6815 (1976).

Tris-HCl

Chelex resin

Ma, C. and Ray, W. J., Biochem., 19, 751 (1980); Yeh, Y.,
et al., Biochem., 18, 882 (1979).

Tris-HCl, DTT

Chelex resin

Miziorko, H. M. and Sealy, R. C., Biochem., 19, 1167 (1980).

Tris-HCl, HEPES-KOH, KCl

Chelex resin

Hunt, J. B. and Ginsburg, A., Biochem., 20, 2226 (1981).

Veronal-buffered saline

Chelex resin

Bartholomew, R. M. and Esser, A. F., Biochem., 19, 2847
(1980).

Water, KOH

Chelex resin

Smithers, G. W. and O’Sullivan, W. J., J. Biol. Chem., 257,
6164 (1982).

Acrylamide

AG 501-X8 resin

Senear, D. F. and Ackers, G. K., Biochem., 29, 6568 (1990).

Formamide

AG 501-X8 resin

Dozin, B., et al., Biochem., 24, 5581 (1985); Jaeger, J. A.,
et al., Biochem., 29, 10147 (1990)

Lubrol-PX

AG 501-X8(D) resin

Moore, A. C., et al., Biochem., 21, 6212 (1982).

Urea

AG 501-X8 resin

Hutchens, T. W. and Yip, T.-T., J. Chromatog., 500, 531(1990); Ropson, I. J., et al., Biochem., 29, 9591 (1990);
Senear, D. F. and Ackers, G. K., Biochem., 29, 6568 (1990).

Endotoxin removal

Affi-Prep
polymyxin support

Talmadge, K. W. and Siebert, C. J., J. Chromatog., 476, 175(1989).

Deionization

5

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Page 6

Protein Concentration and Enrichment
CM Bio-Gel A gel

Weak cationexchanger

Macro-Prep CM
support or Econo-Pac
CM cartridge

Bio-Rex 70 resin
Basic protein

Strong cationexchanger

Macro-Prep S
supports or Econo-Pac
S cartridges

DEAE Bio-Gel A gel
Weak anion
exchanger
Protein sample

Macro-Prep
DEAE support

Acidic protein

Strong anion
exchanger

Macro-Prep Q
supports or Econo-Pac
Q cartridges

AlbuminremovalAffi-Gel blue gel or
Econo-Pac blue
cartridge

Antibody
purification

See Table 1.6

Serum proteins

Bio-Gel HT and
HTP supports or
Econo-Pac HTP
cartridge
Dilute protein
Bio-Gel P-6DG gel

Table 1.4. Guide to Protein Concentration and Enrichment

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Page 7

Protein concentration is often necessary following analysis or purification so thatfurther procedures can be performed. In addition, proteins are generally morestable in relatively concentrated solutions. The optimal method of protein concentration depends on the volume of the sample, the type of protein, the type of
buffer employed, and whether or not the application requires separating the protein from other components.

Dilute Proteins
A rapid and inexpensive method for protein concentration, which eliminates the
problems associated with lyophilization and chemical precipitation, is accomplishedwith Bio-Gel hydroxyapatite media. Hydroxyapatite, a form of calcium phosphate,
has a high capacity for proteins and other macromolecules, yet does not bind low
molecular weight substances. Selective elution with either step or linear phosphate
gradients separates proteins from small molecules and can also concentrate or fractionate proteins. A batch technique, however, provides the most efficient method forconcentrating very dilute protein solutions. Hydroxyapatite media is available either
hydrated or as a dry powder. It is also available in convenient prepacked Econo-Pac
HTP cartridges suitable for small sample preparation applications.
Dilute protein samples can also be concentrated using Bio-Gel P-6DG desalting
gel. The solution to be concentrated is placed in dialysis tubing which is then covered with dry gel. The sample is concentrated as water diffuses out of the tubing
into the gel. Bio-Gel P-6DG gel provides a simple concentration method for samples of 5 ml or greater.

Acidic and Basic Proteins
Ion exchange chromatography can be used to concentrate and purify many proteins.
Since proteins are amphoteric molecules, the net charge of a given protein can be
manipulated by altering its pH. At a pH > pI, proteins are negatively charged, whileat pH < pI, proteins are positively charged (Table 1.5). Thus, cation or anion
exchange resins can be used to bind any given protein. However, since most proteins maintain activity at a physiological pH, it is usually desirable to keep protein
solutions at or near neutral pH. It is also important to consider the pI of the molecule of interest in relation to the pIs of the contaminants to be removed, as it may benecessary to select a pH range outside of the physiological range to obtain thedesired separation.

7

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Page 8

Table 1.5 Properties of Acidic and Basic Proteins at Physiological pH
Protein TypeBinding pH Range

At pH 7Resin TypeAcidic protein
pI < 7

pH = 1 unit > pI

Negative net charge

Anion exchange resin

Basic protein
pI > 7

pH = 1 unit < pI

Positive net charge

Cation exchange resin

Bio-Rad offers strong and weak cation and anion supports for protein concentration and purification. Strong ion exchangers like the Macro-Prep Q and S supports
can be used over a wide pH range (1-10), while weak ion exchangers operate overnarrower pH ranges (see table 1.6). The Macro-Prep supports are available in
Econo-Pac cartridges, which are ideal for small sample preparation applications.
In addition, due to their exceptional mechanical and chemical stability, these supports are ideal for scale-up applications.

Table 1.6 Ion Exchange Supports for Protein Concentration and Purification
SupportType

Functional
LigandOperating
pH Range

Capacity

Macro-Prep Q support
Macro-Prep high Q support

strong anion
strong anion

N+(CH3)3
N+(CH3)3

1-10
1-10

>15 mg BSA/ml>25 mg BSA/ml

Macro-Prep S support
Macro-Prep high S support

strong cationstrong cation

SO3SO3-1-10
1-10>35 mg IgG*/ml>55 mg IgG*/ml

Macro-Prep DEAE support
DEAE Bio-Gel A gel

weak anion
weak anion

N+(C2H5)2
N+(C2H5)2

4-8
2-9.5

>30 mg BSA/ml
45±10 mg/ml
hemoglobin

Macro-Prep CM support
CM Bio-Gel A gel

weak cation
weak cation

COOCOO-

4-13
4.5-10>20 mg BSA/ml
45±10 mg/ml
hemoglobin

Bio-Rex 70 resin

weak cation

COO-

5-14

0.5 meq/ml*Human IgG

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Page 9Serum Proteins
AlbuminAlbumin, the major serum constituent, can be effectively removed from plasma andserum by affinity chromatography with Affi-Gel blue gel. This gel can quickly
remove more than 95% of the serum albumin with little non-specific adsorption of
the other serum proteins. The binding of albumin is so strong that a high concentration of salt or chaotropic reagent is required to desorb it. Other serum proteins
either do not bind to Affi-Gel blue gel or can be eluted with relatively low concentrations of salt. Affi-Gel blue gel is available in two mesh sizes; a faster flowing
50-100 mesh, and a higher capacity, slower flowing 100-200 mesh. It is also available in convenient Econo-Pac blue cartridges for easy sample preparation.

Antibodies

Bio-Rad offers chromatography supports, columns cartridges, and kits for sample
preparation in antibody purification applications. Ion exchange, affinity, gel filtration, hydrophobic interaction (HIC), and hydroxyapatite techniques are all used
for antibody samples. The choice of technique depends upon many factors, including sample complexity, sample volume, and the purification goal. Combinations ofthese techniques can be used to increase the final level of purity (Table 1.7).
Dye-ligand affinity chromatography, using DEAE and CM Affi-Gel blue products,
is useful for the purification of monoclonal and polyclonal antibodies from serum.
CM and DEAE Affi-Gel blue gels are bifunctional gels that combine ion exchange
and dye affinity chromatography to bind albumin and other serum proteins. These
gels will remove albumin and protease from serum or ascites fluid for the preparation of protease-free antibodies or enriched serum samples. DEAE Affi-Gel blue
gel is available in convenient Econo-Pac cartridges, Econo-Pac serum IgG purification columns and in a kit format which includes everything necessary to purify
IgG from serum.
Hydroxyapatite, a crystalline form of calcium phosphate, provides excellent resolution using gentle separation conditions. Hydroxyapatite can be used for the
purification of polyclonal and monoclonal antibodies, the separation of IgG from
IgM, light chain analysis, and the concentration of antibody from large volumes ofcell culture supernatant. Econo-Pac HTP cartridges provide the most convenient
format for using hydroxyapatite with low pressure chromatography set-ups.
Hydroxyapatite chemistry is also available in a Macro-Prep support. The MacroPrep Ceramic Hydroxyapatite support is ideal for scale-up applications due to its
high chemical and mechanical stability.

9

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Page 10

Antibody Sample
Purification of IgG

Immunoaffinity

Protein A
affinity

Ion exchange

Hydroxyapatite

Hydrophobic
interaction

Immobilized
antigencolumnusing
Affi-Gel 10
gelEcono-Pac
protein A
columnsandcartridges

Econo-Pac
Q, S, CM
cartridges

Econo-Pac
HTP
cartridge

Econo-Pac
methylHIC
cartridge

Affi-Gelprotein A
supportsand kits

Macro-Prepionexchangesupports

Bio-Gel andMacro-Prep
Hydroxyapatite
supports

Macro-Prep
HIC
supports

Purify IgM or separate IgG from IgMHydroxyapatite

Ion exchange

Gel filtration

Econo-Pac
HTP
cartridge

Econo-Pac
Q, S, CM
cartridges

Bio-GelA gels

Bio-Gel andMacro-Prep
Hydroxyapatite
supports

Macro-Prepionexchangesupports

Table 1.7. Guide to Antibody Purification
Protein A, a bacterial-coat protein from Staphylococcus aureus, binds the Fc portionof immunoglobulins. This specificity insures a high level of purity. Protein A affinity chromatography is especially useful for the purification of monoclonal mouse
IgG, including mouse IgG1, as well as human and rabbit IgG. Protein A affinity
supports, kits, prepacked columns, and cartridges are available to make theseapplications convenient.

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Page 11

Gel filtration with Bio-Gel A-0.5 ml gel may be used for the separation of IgG fromIgM by exploiting the molecular weight and size differences between them.
Additionally, desalting columns (packed with gel filtration supports) such as
Econo-Pac 10DG columns or Econo-Pac P6 cartridges are useful in many protocols. Desalting or buffer exchange is often required between purification steps andthe use of desalting columns rather than dialysis can save time.
Hydrophobic interaction chromatography (HIC) can be used as a faster and higher
resolution alternative to ammonium sulfate precipitation steps. HIC has beenshown to be the technique of choice for the purification of certain rat antibodies
and as a second step after ion exchange for the purification of mouse antibodies.
Bio-Rad offers a mildly hydrophobic Macro-Prep methyl HIC support and a morehydrophobic Macro-Prep t-butyl support. The Macro-Prep HIC supports are available in 1 ml and 5 ml Econo-Pac cartridges for easy sample preparation.

Econo-Pac cartridges.

11

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Page 12

Table 1.8. Econo-Pac Cartridge Specifications
Application

Cartridge
Name

FunctionalGroup

Protein and plasmid
purification

Econo-Pac-N+(CH3)3
high Q cartridge

5 ml
1 ml

≥170 mg BSA
40 mg BSA

1.0–3.0 ml/min
0.5–1.0 ml/min

Protein and plasmid
purification

Econo-Pac Q
cartridge

5 ml
1 ml

75 mg ferritin
15 mg ferritin

1.0–3.0 ml/min
0.5–1.0 ml/min

Protein purification

Econo-Pac-SO3high S cartridge

5 ml
1 ml

≥230 mg human IgG 1.0–3.0 ml/min
55 mg human IgG
0.5–1.0 ml/min

Protein purification

Econo-Pac S
cartridge

-SO3-

5 ml
1 ml

175 mg human IgG
35 mg human IgG

Protein purification
cartridge

Econo-Pac CM

-COO-

5 ml
1 ml

125 mg hemoglobin 1.0–3.0 ml/min
25 mg hemoglobin 0.5–1.0 ml/min

Protein purification

Econo-Pac
t-Butyl HIC
cartridge-OC(CH3)3

5 ml
1 ml

≥65 mg HSA≥15 mg BSA

0.5–3.0 ml/min
0.5–1.0 ml/min

Protein purification

Econo-Pac
methyl HIC
cartridge

-OCH3

5 ml
1 ml

40 mg BSA
8 mg BSA

0.5–3.0 ml/min
0.5–1.0 ml/min

Protein and nucleic
acid purification

Econo-Pac HTP
cartridge[Ca5(PO4)3OH]2

5 ml

15 mg BSA
30 mg lysozyme
3 mg BSA
6 mg lysozyme

0.5 ml/min

5 ml

34 mg mousemonoclonal IgG
70 mg human IgG

0.5–1.5 ml/min

1 ml

8 mg mousemonoclonal IgG
16 mg human IgG

0.1–0.5 ml/min-N+(CH3)3

Bed
ProteinVolume Capacity

1 ml
Antibody purification

Econo-Pac
protein A
cartridge

Protein A

Recommended
Flow Rate

1.0–3.0 ml/min
0.5–1.0 ml/min

0.6–0.8 ml/min

Desalting, buffer
exchange

Econo-Pac P6
cartridge—

5 ml

100 µl to 3.0 ml
sample volumes

0.5–1.0 ml/min

Purification of
growth factors,
proteases, DNAbinding proteins,endonucleases

Econo-Pac
heparincartridge

Heparin

5 ml

Varies greatly
depending on
the protein

1.0–3.0 ml/min

IgG purification

Econo-Pac DEAE Cibacron Blue
blue cartridge F3GA and DEAE

5 ml

0.3–1.0 ml serum
sample volumes

0.5–2.5 ml/min

Albumin removal,
purification ofserum proteins,various enzymes

Econo-Pac blue Cibacron blue
cartridge
F3GA dye

5 ml

0.3–1.0 ml serum
sample volumes

0.5–2.5 ml/min

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References
Application

Product

ReferenceAlbumin removal from serumAffi-Gel blue gel

Razavi, M.H., et al., Am. J. Hypertens., 1, 91S -95Sm (1988).

Anti-idiotypic antibody
purification

Affi-Gel 10 gel

Lacy, M. J., et al., J. Immunol., 142, 3482(1989).
Lombes, M., et al., J. Immunol., 143, 4078 (1989).

Anti-peptide antibody
purification

Affi-Gelprotein A gel

Okamoto, T., et al., Cell, 62, 709 (1990).

Antibody conjugate purification

Bio-Gel P-30 gel

Zaidi, M., et al., Clin. Chem., 34, 655 (1988).

Antibody coupling

Affi-Gel 10 gel

Brattsand, G., et al., J. Immunol., 144, 3651 (1990).
Douglas, C.M. and Collier, R., J. Biochem., 29, 5043 (1990).

Antibody purification

Bio-Gel A-5m gel

Liu, Z. Y., et al., J. Immunol., 142, 2370 (1989).

Affi-Gel 10 gel

Bequinot, L., et al., Proc. Natl. Acad. Sci. USA, 82, 2774
(1985); Feeney, A. J. and Thueraur, D. J., J. Immunol., 143,
4061, (1989); Larrodera, P., et al., Cell, 61, 1113 (1990);
Seqev, N., et al., Proc. Natl. Acad. Sci, USA, 82, 1531 (1985).

Affi-Gel 15 gel

Makker, S. P., et al., J. Immunol., 142, 2264 (1989).

Protein A MAPS
buffers

Tomkinson, B.E., et al., J. Immunol., 142, 2230 (1989).

Antibody purification fromcrude antiserumAffi-Gel 15 gel

Wildgoose, P., et al., Biochem., 29, 3413 (1990).

Antigen purification

Affi-Gel 10 gel

Brooks, K. H. and Feldbush, T. L., J. Immunol., 127, 963(1981); Suzuki, Y., et al., J. Clin. Microbiol., 28, 1734
(1990).

Antigenic protein fractionation

Hydroxyapatite

Vosti, K. L., J. Med. Microbiol., 11, 453 (1978).

Antisera purification

Affi-Gel 10 gel

Glick, A.B., et al., Cell Regulation, 1, 87 (1989).

Affi-Gel 15 gel

Enos, A. P., and Morris, N. R. , Cell, 60, 1019 (1990).

Ascites fluid absorption

Affi-Gel blue gel

Lacy, M. J., et al., J. Immunol., 142, 3482 (1989).

DNA A protein separationfrom E. coli

Bio-Rex 70 resin

Sekimizu, K., et al., J. Biol. Chem., 263, 7136 (1988).

Enzyme purification

Hydroxyapatite

Barry, III, C. E., et al., Biochem., 28, 6323 (1989); Schott, K.,
et al., J. Biol. Chem., 265, 4204 (1990); Ghosh, G., et al.,Biochem., 29, 2220 (1990); Shayiq, R. M. and Avadhani, N.
G., Biochem., 29, 866 (1990).

Exonuclease purification

Bio-Rex 70 resin

Perrino, F. W. and Loeb, L. A., Biochem., 29, 5226 (1990).

Hydroxyapatite

Perrino, F. W. and Loeb, L. A., Biochem., 29, 5226 (1990).

Extraction of GIIIA

Bio-Rex 70 resin

Cruz, L. J., et al., Biochem., 28, 3437 (1989).

F(ab)2 fragment purification

Affi-Gelprotein A gel

Tsuchiya, N., et al., J. Immunol., 144, 4742 (1990).

13

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Page 14

Application

Product

Reference

Factor D purification

Hydroxyapatite

Fry, M., et al., Biochem., 24, 7549 (1985).

Fractionation of plasma proteinsAffi-Gel blue gel

Gianazza, E. and Arnaud, P., J. Biochem., 201, 129 (1982).

Glycoprotein purification

Hydroxyapatite

Gorbunoff, M. J., J. Chromatog., 187, 224 (1980).

Hemocyanin purification

Bio-Rex 70 resin

Moore, M. D., et al., J. Biol. Chem., 261, 10511 (1986).

Hemoglobin separationfrom blood

Bio-Rex 70 resin

Ersser, R. S., et al., Biomedical Chromatog., 1, 183 (1986).

Histone purification

Bio-Rex 70 resin

D’Anna, J. A., et al., Biochemistry, 18, 943 (1979): Walker,J., et al., J. Biol. Chem., 265, 5736 (1990).

IgG purification

DEAE Affi-Gel
blue gelHori, M. T. and Abrass, C. K., J. Immunol., 144, 3849
(1990).

Affi-Gelprotein A gel

Moellering, B. J., et al., BioPharm, 30 (Feb. 1990).

Bio-Gel A-1.5m gel

Graham, I. L., et al., J. Immunol., 142, 2352 (1989).

DEAE Affi-Gel
blue gelAndrews, R. K., et al., Biochem., 28, 8326 (1989);
Usui, S., et al., Biochem., 29, 4618 (1990).

IgG quantitation

Affi-Prep
protein A gel

Compton, B. J., et al., Anal. Chem, 61, 1314 (1989).

IgG removal

Affi-Gel 10 gel

Alters, S. E., et al., J. Immunol., 1454, 4587 (1990).

IgG1 coupling

Affi-Gel 10 gel

Tawil, N. J., et al., Biochem., 29, 6540 (1990).

IgG1 purification

Affi-Gel blue gel

Shen, L., et al., J. Immunol., 143, 4117 (1989); Tawil, N.J.,
et al., Biochem, 29, 6540 (1990).

IgG1, IgG2a and IgM purification

Affi-Gelprotein A column
and hydroxyapatite

Altieri, D. C. and Edgington, T. S., J. Immunol., 145,
246 (1990).

IgM and IgG purification fromsheep and cattle serum

CM Affi-Gel
blue gel

Schmerr, M. J. F., et al., J. Chromatog., 326, 225 (1985).

IgM purification from serum

Bio-Gel P-200 gel

Pretzman, et al., J. Immunol. Methods, 83, 301 (1985).

Immune complex isolation

Affi-Gelprotein A gel

Nagafuchi, A. and Takeichi, M., Cell Regulation, 1, 37(1989).

Bio-GelA-1.5m gel

Levinson, S. S., et al., Clin. Chem., 34, 784 (1988).

Isolation of human albumin

CM Affi-Gel
blue gel

Podulso J. F. and Curran, G. L., Proc. Natl. Acad. Sci. USA,89, 2218 (1992).

Lambda and Kappa lightchain separation

DEAE Bio-Gel A gel

Dalal, F. R. and Winsten. S., Clin. Chem., 25, 1 (1979).

Lipoprotein fractionation

Hydroxyapatite

Kostner, G. and Holasek, A., Biochim. Biophys. Acta, 488,
417 (1977).

Monoclonal antibody coupling

Affi-Gel 10 gel

Wilden, P.A., et al., Biochem., 28, 9734 (1989).

14

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Page 15

Application

Product

Reference

Monoclonal antibodies—separation of different classes

Hydroxyapatite

Mahoney, C. W. Q., et al., J. Biol. Chem., 2265, 5424(1990).

Monoclonal antibody affinity
chromatography

Affi-Gel 10 gel

Hammerberg, B., et al., J. Immunol., 143, 42901(1989).

Monoclonal antibody
purification

Affi-Gel 10 gel

Hsu, Y., et al., Proc. Natl. Acad. Sci. USA, 81, 2107
(1984); Theodos, C. M., et al., J. Immunol., 144, 4011
(1990).

Affi-Gelprotein A column

Dueweke, T. J. and Gennis, R. B., J. Biol. Chem., 265, 4273
(1990).

Protein A MAPS
buffers

Gogol, E. P., et al., Biochem., 28, 4717 (1989).

DEAE Affi-Gel
blue gelFung, S. K-K, et al., Biochem, 29, 2657 (1990); Bruck, C.,et al., Methods Enzymol., 121, 587-596 (1986).

Monoclonal antibody
purification from ascites

DEAE Affi-Gel
blue gel

Wildgoose, P., et al., Biochem., 29, 3413 (1990).

Muscle protein separation

Hydroxyapatite

Suzuki, A., et al., J. Biol. Chem., 251, 6860 (1976).

Nucleic acid basecomposition determination

Hydroxyapatite

Miyazawa, Y. and Thomas, Jr., C. A., Mol. Biol., 11, 223(1965).

Peptide purification

Bio-Rex 70 resin

Wilson, S. P., J. Neurosci. Methods, 15, 155 (1985).

Phosphoprotein separation

Hydroxyapatite

Addeo, F., et al., J. Dairy Res., 44, 63 (1977); Donnelly, W.
J., J. Dairy Res., 44, 63 (1977).

Polypeptide purification from
Croatalus atrox venom

Bio-Rex 70 resin

Hamilton, S. L. et.al., Science, 229, 182 (1985).

Protein concentration

Hydroxyapatite

Tiselius, A., et al., Arch. Biochem. Biophys., 65, 132 (1956).

Rhodamine removal fromrhodamine-conjugatedantibodies

Bio-Beads SM-2 beads

Spack, et al., Anal. Biochem., 158, 233 (1986).

Topoisomerase I purification

Hydroxyapatite

Hertzberg, R. P., et al., Biochem., 28, 4629 (1989).

Topoisomerase purification

Hydroxyapatite

Saijo, M., et al., Biochem., 29, 583 (1990).

15

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Page 16

16

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Page 17

Desalting and Buffer Exchange
MW 6,000 - 40,000
or 5 bp nucleic acid

Bio-Spin 6
columns

MW > 40,000
or 20 bp nucleic acid

Bio-Spin 30
columns

Volume < 0.1 ml

Volume 1 - 3 ml
Econo-Pac
10DG column
MW > 2,000
MW > 6,000

Volume 0.1 - 3 ml
Econo-Pac P6
cartridge

Volume > 0.1 ml

SampleAmino acids andpeptidesMW < 2,000

Volume > 3 ml
Bio-Gel
P-6DG gel

MW 4,000 - 6,000

Bio-Gel P-4 gel

MW 2,000 - 4,000

Bio-Gel P-2 gel

AG 11 A8 resin
AG 50W-X8 resin

Hydrophobic sample

Bio-Beads SM
adsorbents

Peptide with tyrosine
or tryptophan residue

Bio-Beads SM
adsorbents

Table 2.1. Guide to Desalting and Buffer Exchange

Introduction
Desalting and buffer exchange are often required in order to perform many
analytical assays or for the next step in a purification scheme. Dialysis, the mostcommon technique for desalting and buffer exchange can be very time consuming.

17

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Page 18

A more desirable alternative for desalting and buffer exchange is gel filtration.
This method is as effective as exhaustive dialysis in far less time. Other desalting
and buffer exchange methods include ion retardation, adsorption, and ion
exchange chromatography. The method to be used for salt removal or buffer
exchange depends on the properties of the sample, its molecular weight, and thevolume to be desalted. Table 2.1 can help you select among the products available
for desalting and buffer exchange.

Methods
MW > 2,000 Daltons (Gel Filtration)
Desalting and buffer exchange of samples with molecular weights greater than
2,000 daltons is easily accomplished using Bio-Gel P polyacrylamide gel filtration
media. In gel filtration chromatography, small molecules enter the pores of the gel
and are retained, while large molecules are excluded from the pores and eluted in
the void volume. In this way salts, including buffer salts, are separated from larger
molecules such as proteins and nucleic acids. Bio-Gel P gels are available inseveral particle size ranges with exclusion limits ranging from 1,800 to 100,000
daltons. Bio-Gel P-2, P-4, and P-6 gels are excellent for removing salts fromsamples containing biomolecules larger than 1,800, 4,000, and 6,000 daltons,
respectively. Bio-Gel P gels are available in bulk for self-packing or in convenientprepacked columns and cartridges. Bio-Spin 6 and 30 columns are recommended
for small sample volumes (< 100 µl). These columns combine gel filtration andcentrifugation to provide rapid, convenient desalting while minimizing sample
dilution. For samples larger than 100 µl, self-packed or prepacked Econo-Pac
10DG columns and Econo-Pac P6 cartridges are recommended. These columns
can desalt samples up to 3 ml in minutes (Figure 2.1 and 2.2).

MW < 2,000 Daltons (Ion Exchange and Adsorption)
Ion retardation, ion exchange, and adsorption chromatography are extremely
useful techniques for desalting and buffer exchange applications when the biomolecules to be desalted are smaller than 2,000-4,000 daltons. Small amphotericmolecules, such as amino acids and peptides, can be desalted using AG 11 A8 ion
retardation resin or AG 50W-X8 cation exchange resin. AG 11 A8 resin is used fordesalting and buffer exchange of amphoteric compounds up to 4,000 daltons,
while AG 50W-X8 ion exchange resin is used for desalting cationic compounds up

18

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Page 19

to a molecular weight of 1,000 daltons. AG 50W-X8 resin is especially useful foramino acid samples, and is the resin of choice when the sample is in acetate or citrate buffer. AG 50W-X8 resin should not be used for hydrophobic compounds
because they will non-specifically adsorb to the resin matrix. AG 50W-X8 resin isavailable in several particle size ranges and in prefilled Poly-Prep columns foradded convenience. Bio-Beads SM adsorbents are ideal for desalting hydrophobic
compounds and are recommended for desalting peptides containing tyrosine or
tryptophan residues.
Conditions
Column:Econo-Pac 10DG
desalting column
Sample:Bovine serum albumin in
250 mM NaCl, 3.0 ml
Eluant:H2O
Flow rate:
Peaks:

1.0 ml/min
1. BSA,
2. NaCl

1122

Conductivity
Conductivity
O.D.280
280
O.D.

Fig. 2.1. Protein desalting with the Econo-Pac 10DG desalting column.

11

Conditions
Column:Econo-Pac 10DG
desalting column
125I labeled follicle stimulating
Sample:hormone, 3.0 ml
Eluant:H2O
Flow rate:
1.0 ml/min
Peaks:
1. 125I labeled FSH,
2. 125I (unincorporated)

22

Fig. 2.2. Separation of radiolabeled protein from unincorporated radiolabel.

19

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Page 20

References
Application

Product

Reference

Desalting amino acids

AG 50W-X8 cationexchange resin

Sato, M. and Yagi, K., J. Chromatog., 242, 185 (1982).

Desalting amino acids
and peptides

AG 11 A8 ion
retardation resin

Heathcote, J. G., et al., Clin. Chem. Acta., 32, 457 (1971).

Desalting

AG 1-X4 resin

Turco, S. J. and Pickard, J. L., J. Biol. Chem., 257, 8674
(1982).

AG 50W-X2 resin

Nakamura, T., et al., J. Biol. Chem., 265, 5390 (1990).

AG 50W-X4 resin

Esmon, B., et al., J. Biol. Chem., 259, 10322 (1984).

Bio-Gel P-2 gelFaure, G., et al., Biochem., 22, 2068 (1983);. Gariepy, J. and
Hodges, R. S., Biochem., 22, 1586 (1983); Gatineau, E.,
et al., Biochem., 29, 6480 (1990); Kehry, M. R., et al.,Biochem., 21, 5415 (1982); Knibbs, R. N., et al., Biochem.,
28, 6379 (1989); Loganathan, D., et al., Biochem., 29, 4362
(1990); McKinney, J. D., et al., Anal. Chem., 55, 91 (1983);
Murakami, K., et al., Biochem., 21, 5488 (1982); Matsumiya,
M. and Otake, S., Bull. Jpn. Soc Fish, 52, 1617 (1986); Van
Pelt, J. E. and Northrop, D. B., Arch. Biochem. Biophys., 230,
250 (1984).

Bio-Gel P-6 gel

Walsh, M. T., et al., Biochem., 29, 6 (1990); Keller, J. W.,
et al., J. Biol. Chem., 265, 5531 (1990).

20

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Plasmid Purification and Probe Clean-up
Cellularcontaminants
PCR-inhibiting
contaminants

Prep-A-Genematrix or kit
InstaGene kit
Chelex 100 resin

Volume > 3 ml
Bio-Gel P-6DG gel

Contaminants< 6,000 or 5 bp
nucleic acid

Volume 0.1-3 ml
Econo-Pac 10DG
column or EconoPac P6 cartridge
Volume < 100 µl
Bio-Spin 6 column

Nucleic acidsampleNucleotides orother ioniccontaminantsContaminants< 40,000 or 20 bp
nucleic acidContaminants< 60,000 or 55 bp
nucleic acid
DNA isolationfrom agarose gel

Ethidium bromide
removalVolume > 100 µl
Bio-Gel 30 gel
Volume ≤ 100 µl
Bio-Spin P-30
columnBio-Gel P-60 gel

Prep-A-Genematrix or kit
Molecular Biology
Grade AG 50W-X8
resin
Bio-Beads SM
adsorbents

Table 3.1. Guide to Plasmid Purification and Probe Clean-up

Introduction
It is often necessary to separate plasmid DNA from cell lysates or compounds suchas unincorporated nucleotides, unincorporated linkers, and ethidium bromide, orto remove radiolabel or perform buffer exchange on nucleic acids and oligonucleotides. Table 3.1 can help you select among the products available for plasmid,
oligonucleotide, and probe purification.

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Page 22

Applications
Ethidium Bromide and Propidium Iodide Removal
The intercalating dye reagents, ethidium bromide and propidium iodide, are commonly used to visualize single- and double-stranded nucleic acids. The presence of
these dyes may inhibit restriction enzyme function, transformation efficiencies,
and other applications utilizing DNA and RNA. AG 50W-X8 ion exchange resin is
useful for removing both ethidium bromide and propidium iodide after use. The
fluorescent labeled nucleic acid is simply passed over a column of the AG 50W-X8
resin, which removes the dye, and the pure preparation is collected in the void
volume. Alternatively, ethidium bromide can be removed from nucleic acid preparations using Bio-Beads SM adsorbents.

DNA Isolation from Agarose Gel
The Prep-A-Gene DNA purification matrix provides an easy, efficient method forrecovering DNA from agarose gels. The novel matrix can be used to extract,
purify, and concentrate DNA in less than 20 minutes. In a simple procedure, DNA
is bound to the matrix while contaminants are washed away. The DNA is subsequently eluted from the matrix, yielding recoveries in excess of 85%. The purified
DNA can be used for transformation, restriction enzyme digestion, ligation, andsequencing procedures. The Prep-A-Gene matrix is available separately or in convenient Prep-A-Gene DNA purification kits which include the binding, wash, andelution buffers.

Radioactive Label Removal
The removal of unincorporated radioactive label is often required in nucleic acidexperiments or is necessary to prevent interference in subsequent assays. For
example, unincorporated radioactive label removal is very important to determine
the labeling efficiency of a labeling reaction. Also DNA free of unincorporated label
is required when the DNA is to be used as markers for gel electrophoresis, as
substrates for enzymatic reactions, or as probes for hybridization. Bio-Rad offers
several alternatives for radiolabel removal including gel filtration and adsorption
chromatography. Gel filtration chromatography using Bio-Spin columns is ideal
for removing unincorporated nucleotides. Separation occurs as the small nucleotide
molecules are retained in the pores of the gel while the larger DNA molecules
are eluted in the elution volume. These prepacked columns are quality-control

22

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Page 23

certified to insure a high recovery of DNA and a high retention of unincorporated
nucleotides. Bio-Spin 6 columns are suitable for the purification of nucleic acids
greater than ~ 5 bp in length, while Bio-Spin 30 columns are best suited for nucleic
acids greater than ~ 20 bp in length. Bio-Gel P gel is also available in bulk,
prepacked cartridges, and Econo-Pac 10DG columns for larger sample volumes.
Alternatively, the Prep-A-Gene DNA purification kit efficiently removes nucleoside
triphosphates from radiolabeling reactions. DNA binds to the matrix while theunincorporated nucleotide triphosphates do not. The unincorporated label iswashed away with other reaction components and the DNA is subsequently eluted
in a purified, concentrated form. Finally, nucleotides can also be concentratedusing Bio-Beads SM-2 or SM-7 adsorbents.
End cap

Fully autoclavable

Graduated volume
markings
Collection tube adaptor
ring

Porous polymer bed
supportLuer column tip,with snap off tip

Bio-Spin columns.

Plasmid Purification
Methods for plasmid purification include adsorption, size exclusion, and ion
exchange chromatography. Bio-Rad offers the following products forplasmid purification.

23

8/11/98 7:23 AM

Page 24Prep-A-Gene DNA or plasmid purification kits provide a fast and efficient means
for DNA purification. The kits provide everything necessary for the purificationand concentration of DNA > 200 bp. Using a batch technique, DNA is adsorbed
onto the silica-like matrix, and RNA, protein, and other cellular components are
washed away. The purified DNA is then eluted using elution buffer and is recovered in a form immediately available for use in cloning protocols. Using the PrepA-Gene matrix and the GS Gene Prep manifold, 24 single-strand DNA (M13,
phagemid) samples can be purified for nucleotide sequence analysis from phagesupernatants in 45 minutes. Up to 24 double-strand DNA (plasmid) samples canbe purified for either restriction enzyme analysis or nucleotide sequence analysis
from cleared bacterial lysates in 2 hours.
Ion exchange chromatography is yet another method for plasmid purification.
Plasmids can be purified using the Macro-Prep Q support without the need for
cesium chloride gradients or phenol/chloroform extractions. This method produces
plasmid DNA of the purity necessary for restriction enzyme digests, ligation reactions, and transformation protocols (Figures 3.1 and 3.2). The Macro-Prep Q support is available in convenient Econo-Pac cartridges in both 1 ml and 5 ml sizes.

Detection:

2

3

4

5

Purified plasmid

Flow rate:
Cartridge:

1

Start gradient

Gradient:

10 ml lysate from 1 liter overnight culture
Buffer A:
50 mM MOPS, pH 8.0
15% ethanol
0.3 M NaCl
Buffer B:
50 mM MOPS, pH 8.0
15% ethanol
0.75 M NaCl
0-100% B
in 40 min
1.5 ml/min
Econo-Pac Q
cartridge, 5 ml
254 nm,2.56 AUFS
Injection 1
Injection 2
Injection 3
Injection 4

Sample:Conditions:A254

D SPG Pt. 3

Time

Lane 1: pUC19 uncut
Lane 2: pUC19 EcoR I
Lane 3: pUC19 BamH I

100 minFig. 3.1. Purification of pUC19 using Econo-Pac
Q cartridge.

24

Lane 4: pUC19 Hae II
Lane 5: l HindIII

Fig. 3.2. Verification of plasmid purity. 1%
agarose gel of 0.5 ng pUC 19 purified with
Econo-Pac Q cartridge.

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Page 25

Removal of PCR Inhibiting Contaminants
The InstaGene kit provides convenience and dependability in removing
PCR-inhibiting components from genomic DNA preparations prior to PCR amplification. The unique matrix binds contaminants of the PCR procedure while the
DNA remains in solution. A simplified protocol eliminates phenol/chloroform
extractions and deproteinization steps. The kit is functionally tested to insure its
ability to provide a useful DNA template for PCR amplification. Biotechnology
grade Chelex 100 resin can also be used in DNA preparations prior to PCR amplification, however, the resin is not functionally tested for this application.

References
Application

Product

Reference

cDNA size fractionation

Bio-Gel A-50m gel

Giese, K. and Subramanian, A. R., Biochem., 28, 3525
(1989).

Chromatin fragment purification

Bio-Gel A-50m gel

Jin, Y. J. and Cole, R. D., J. Biol. Chem., 261, 3420 (1986).

Chromatin isolation

Bio-Gel A-0.5m gel

Prevelige, Jr., P. E. and Fasman, G. D., Biochem., 26, 2944
(1987).

DNA fragment purification

Bio-Gel P-60 gel

Schmidel, D. K., et al., Biochem., 29, 7845 (1990).

DNA preparation for PCR
amplification

Chelex 100 resin

Walsh, P. S., et al., BioTechniques, 10, 506 (1991).

Ethidium bromide removal

AG 50W-X8 resin

Maniatis, T., Fritsch, E.F.and Sambrook, J. Molecular
Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratory (1982).

Bio-Beads SM
adsorbents

Joshua, H., BioTechniques, 4, 207 (1986); Lunn, G. andSansone, E. B., Biotechnic and Histochemistry, 66, 307(1991).

Isolation and purification
plasmid DNA

Bio-Gel A gel

Rachubinski, R. A., et al., Proc. Nat. Acad. Sci. USA, 82, of
3973 (1985).

Labeled DNA recovery

Bio-Gel P-60 gel

Shoubridge, E. A., et al., Cell, 62, 43 (1990).

Labeled RNA
separated from ATP

Bio-Gel P-2 gel

Blum, B. and Simpson, L., Cell, 62, 391 (1990).

Linear duplex
DNA sample extraction

Bio-Gel A-15m gel

Colowick, S. P. and Kaplan, N. O., Meth. Enzymol., 68, 43(1980).

Oligomer desalting

Bio-Gel P-2 gel

Krug, M., et al., Biochem., 21, 4713 (1982).

Oligomer fractionation

Bio-Gel P-2 gel

Keller, K. M., et al., Biochem., 28, 8100 (1989).

Oligonucleotide clean-up

Bio-Gel P-6DG gel

England, T. E. and Uhlenbeck, O. C., Biochem., 17, 2069
(1978).

25

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Page 26

Application

Product

Reference

Oligonucleotide desalting

Bio-Gel P-100 gel

Hogrefe, H. H., et al., J. Biol. Chem., 265, 5561 (1990).

Econo-Pac 10DG
desalting columns

Fischer, I., et al., BioTechniques, 9, 300 (1990).

Bio-Gel P-60 gel

Conner, G. E., et al., Biochem., 28, 3530 (1989).

Bio-Gel P-6DG gel

Ramsing, N. B., et al., Biochem., 28, 9528 (1989).

Chelex resin

Kochoyan, M., et al., Biochem., 29, 4799 (1990).

Hydroxyapatite

Grohmann, K., et al., Biochem., 14, 1961 (1975).

Plasmid purification

Prep-A-Gene DNA
purification kit

Willis, E. H., BioTechniques, 9, 92 (1990)

Propidium iodide removal

AG 50W-X8 resin

Rodrigues, R. L.,and Tait, R. C., Recombinant DNA
Techniques: An Introduction, Addison-Wesley PublishingCo., Reading, Massachusetts, 153-158 (1983).

Bio-Beads
SM adsorbents

Lunn, G. and Sansone, E. B., Biotechnic and Histochemistry,
66, 307 (1991).

Oligonucleotide purification

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Detergent Removal

DiluteproteinsamplesSensitive
proteinsamples

Samplewithdetergent

Bio-Gel HT
or HTPsupports
Econo-Pac
HTP
cartridge
Bio-Gel HT
or HTPsupports
Econo-Pac
HTP
cartridge

AG 50W-X8
resin
Cationic
detergent

Ionic
detergents
Mostproteinsamples

Anionic
detergent
SDS
Non-ionic
detergent
Triton
X-100

Hydrophobic
proteinsamples

Bio-Beads
SM-2
adsorbents

Bio-Beads
SM-2
adsorbents
AG 11 A8
resin orAG 1-X2
resin
Bio-Beads
SM-2
adsorbents

Bio-Gel HT
or HTPsupports
Econo-Pac
HTP
cartridge

Table 4.1. Guide to Detergent Removal

Introduction
Detergents are used to solubilize proteins in many purification protocols and analytical assays. It then becomes necessary to remove the detergents due to theirinterference with subsequent procedures. In some cases, detergents interfere withprotein analysis and quantitation. Detergent molecules are amphiphilic molecules
consisting of a hydrophobic “tail” and a hydrophilic “head”. The hydrophilic headmay contain either ionic or nonionic chemical structures. The type of detergent
and the stability or sensitivity of the protein present in the sample determines the
optimal detergent removal method.

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Methods
Non-Ionic DetergentsHydrophobic interaction chromatography with Bio-Beads SM-2 beads is ideal for
removing non-ionic detergents such as Triton X-100, NP-40, Tween-20, and octylglucoside. The hydrophobic beads adsorb the hydrophobic tail of the detergent foreasy removal. Removal can be accomplished by either batch or column methods
and the beads can be used in aqueous solutions or with a variety of solvents.

Ionic Detergents
Ion exchange chromatography is recommended for the removal of ionic detergents. Ion exchange resins are available to remove either cationic or anionicdetergents. AG 11 A8 ion retardation resin provides a rapid and efficient method
for removing anionic detergents such as SDS from proteins. AG 50W-X8 strong
cation exchange resin is recommended for removing cationic detergents from protein samples. Alternatively, Bio-Beads SM-2 adsorbents can be used to remove
ionic detergents from small sample volumes. As with non-ionic detergent removal,
the hydrophobic tail of the ionic detergent is adsorbed by the hydrophobic beads.

Sensitive or Dilute Protein Samples
Hydroxyapatite is recommended for removing detergents from samples containing
sensitive enzymes or other proteins which may be affected by exposure to ion
exchange resins. Hydroxyapatite is particularly useful with dilute protein solutions. Protein binds to the hydroxyapatite support, while the detergent isunretained. The protein is subsequently eluted from the column.

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References
Application

Product

Reference

Detergent dialysis

Bio-Beads
SM-2 beads

Wang, C-Y. and Huang, L., Biochem., 28, 9508 (1989).

Detergent removal

Bio-Beads
SM-2 beads

Ator, M. A., et al., Biochem., 28, 9633 (1989); Shayiq, R. M.
and Avadhani, N. G., Biochem., 29, 866 (1990)

Hydroxyapatite

Marcus, C. B., et al., Biochem., 24, 5115 (1985).

Detergent removal, NP-40

Bio-Beads
SM-2 beads

Momoi, T., Biochem. Biophys. Res. Commun., 87, 541 (1979);
Verkman, A. S., et al., Biochem., 28, 4240 (1989).

Detergent removal, C12E8

Bio-Beads
SM-2 beads

Levy, D., et al., Biochem., 29, 9480 (1990).

Detergent removal, cholate

Bio-Beads
SM-2 beads

Bonomo, E. A. and Swaney, J. B., J. Lipid Research, 29, 380(1988); Bonomo, E. A. and Swaney, J. B., Biochem., 29, 5094
(1990); Jinks, D. C. and McElhaney, R. N., Anal. Biochem.,
164, 331 (1987).

Detergent removal, deoxycholate

Bio-Beads
SM-2 beads

Horigome, T. and Sugano, H., Anal. Biochem., 130, 393(1983); Lorusso, D. J. and Green, F. A., Science, 188, 66(1974); Shechter, I. and Bloch, K., J. Biol. Chem., 246, 7690
(1971); Garland, R. C. and Cori, C. F., Biochem., 11, 4712
(1972).

Detergent removal, Emulgen 911

Bio-Beads
SM-2 beads

Gibson, G. G. and Schenkman, F. B., J. Biol. Chem., 253,
5957 (1978); Warner, M., J. Biol. Chem., 257, 12995(1982).

Detergent removal, Emulgen 913

Hydroxyapatite

Nguyen, L. B., et al., J. Biol. Chem., 265, 4541 (1990).

Bio-Beads
SM-2 beads

Brunch, R. C., J. Biol. Chem., 261, 9450 (1986).

Bio-Beads
SM-2 beads

Novick, S. L. and Baldeschwieler, J. D., Biochem., 27, 7919
(1988).

AG 11 A8 resin

Young, D. H., et al., Appl. Environ. Microbiol., 50, 605(1985).

AG 1-X2 resin

Weber, K. and Kuter, D. J., J. Biol. Chem., 246, 4504 (1971).

Bio-Gel P-4 gel

Millett, F., et al., Biochem., 22, 546 (1983).

SDS from proteins

AG 11 A8 resin

Kapp, O. H. and Vinogradov, S. N., Anal. Biochem., 91, 230(1978).

Detergent removal,
sodium cholate

Hydroxyapatite

Nguyen, L. B., et al., J. Biol. Chem., 265, 4541 (1990).

Detergent removal,
Emulphogene BC-720
Detergent removal,
Octylglucoside
Detergent removal, SDS

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Application

Product

Reference

Detergent removal, Triton X-100

Bio-Beads
SM-2 beads

Bonomi, F. and Kurtz, D., Anal. Biochem., 142, 226 (1984);
Bonomo, E. A. and Swaney, J. B., J. Lipid Research, 29, 380(1988); Braun, P., et al., Biochem., 29, 10376 (1990);
Drexler, G., et al., J. Immunol. Methods, 95, 117 (1986);
Holloway, P. W., Anal. Chem., 53, 304 (1973); Metsikko, K.,
et al., EMBO Journal, 5, 3429 (1986); Welling, G. W., et al.,J. Chromatog., 297, 101 (1984).

Detergent removal, Tween-20

Bio-Beads
SM-2 beads

Lamphear, B. J. and Panniers, R., J. Biol. Chem., 265, 5333
(1990).

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Ionic Contaminant Removal
Introduction
Ionic contaminant removal is an important sample preparation step prior to analysis or purification of compounds. Applications vary from general protocols forbuffer deionization and metal ion removal to unique applications for removal of
specific ionic contaminants. Bio-Rad products for ionic contaminant removal
include anion exchange resins, cation exchange resins, mixed bed resins, and ion
retardation resins.

Deionization
Volume > 1 liter
AG 501-X8 resin
using column method
Reagents, buffers,
and water
Volume < 1 liter
AG 501-X8 resin
using batch method

AG 501-X8 resin
using batch method
Deionization

Carbohydrate sample
AG 2 or AG 4 resin
followed by AG 50W resin

AG 501-X8 resinwith dialysis tubing
Protein sample
AG 11 A8 resin

Table 5.1. Guide to Deionization
Typical deionization applications include deionization of carbohydrates, separation of unincorporated label from a labeled sample, removal of ionic contaminantsfrom protein solutions, and deionization of other reagents such as water, formamide, acrylamide, urea, and glyoxal. Table 5.1 lists products for specificdeionization applications.

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Mechanism
AG 501-X8 mixed bed resins are specifically designed for deionization protocols.
Mixed bed resins consist of equivalent amounts of strong cation and strong anion
exchange resins, in their respective hydrogen and hydroxyl forms. Deionization isachieved by exchanging solute cations for hydrogen and solute anions forhydroxyl, with the resulting neutralization yielding water.
Resin-H + Resin-OH + NaCl → Resin-Na + Resin-Cl + H2O
The separation mechanism of the AG 11 A8 resin is ionic adsorption rather thanconventional ion exchange. The AG 11 A8 resin contains strong anion exchangegroups and weakly acidic cation exchange groups which retard ions as they passthrough a column. Adsorbed salts move down the column during water elution
because the fixed exchange groups compete with the mobile salt ions to become
self-adsorbed.

Methods
Deionization may be achieved by either batch or column methods. With the batch
method, the resin is added directly into the solution to be deionized, followed bystirring to achieve deionization. The column method requires preparing a chromatography column and pouring the solution to be deionized over the column.
The results shown in Table 5.2 demonstrate that the column method provides
higher purity in less time. However, the batch method is more commonly used,since it provides reagents of acceptable quality and is more convenient.

Table 5.2. Batch vs Column Deionization of Urea with Mixed Bed Resin
Batch

ColumnSample

100 ml 6 M urea

100 ml 6 M urea

Starting conductivity

70 µmho/cm

70 µmho/cm

Amount of AG 501-X8 resin

5g

Final conductivity

5.0 µmho/cm

Time

5 hours32

5g
0.2 µmho/cm
10 minutes

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Applications
Reagents, buffers,
and water

Reagent preparation applications include deionization of water and other non-ionicsolutions. AG 501-X8 mixed bed resins are recommended for complete deionization of non-electrolyte solutions. The batch method provides a convenient way todeionize electrophoresis reagents. The procedure was originally described byManiatis1 for deionization of formamide, but can be used for other non-ionic
reagents such as urea, glyoxal, and acrylamide. Generally, for reagent preparation
applications, the batch method is recommended for sample sizes of less than 1liter and the column method is recommended for sample sizes in excess of 1 liter.

Protein samples

Protein samples can be deionized using AG 501-X8 mixed bed resins. However,care must be taken to eliminate non-specific binding of hydrophobic proteins to
the resin matrix. Non-specific binding of protein is of special concern when protein concentrations are low or when protein samples are very valuable. Non-specific binding may be eliminated by placing the mixed bed resin in dialysis tubing
to prevent direct contact between the protein and the resin.

Carbohydrate
samples

Carbohydrate samples are also deionized using AG 501-X8 mixed bed resins. Nonspecific adsorption of carbohydrates and polyhydric alcohols can be minimized byusing a batch technique and removing the resin from the carbohydrate solution
immediately following deionization. Alternatively, tandem columns of AG 2-X8
and AG 50W-X8 resin can be used to remove ions from carbohydrates, dextrans,
and polyhydric alcohols. Additionally, sucrose, fructose, and glucose are successfully deionized with AG 4-X4 and AG 50W-X8 resins.

References
Application

Product

Reference

Deionization

AG 50W-X2 resinde Groot, H. J. M., et al., Biochem., 29, 6873 (1990).

AG 50W-X8 resin

Engelhard, M., et al., Biochem., 28, 5432 (1989).

Acrylamide

AG 501-X8 resin

Senear, D. F. and Ackers, G. K., Biochem., 29, 6568 (1990).

Carbohydrates

AG 2-X8 resin

Cullen, M. P., et al., J. Chromatog., 337, 29 (1985).

Formamide

AG 501-X8 resin

Boime, I., et al., J. Biol. Chem., 251, 820 (1976); Dozin, B., etal., Biochem., 24, 5581 (1985); Jaeger, J. A., et al., Biochem.,
29, 10147 (1990); Maniatis, et al., Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Lab (1982);
Nudel, U., et al., J. Biol. Chem., 252, 2182 (1977); Ofengand,J. and Liou, R., Biochem., 19, 4814 (1980).

Lubrol-PX buffer

AG 501-X8(D) resin

Miller, J. A., et al., Biochem., 22, 462 (1983); Moore, A. C.,
et al., Biochem., 21, 6212 (1982).

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Application

Product

Reference

Membrane

Dowex 50 resin

Dunach, M., et al., Biochem., 28, 8940 (1989).

PeptideDowex 1 resin

Palczewski, K., et al., Biochem., 28, 8764 (1989).

Urea

AG 1-X8 resin

Nelson, D. A., et al., Biochem., 21, 4350 (1982); Senear, D.
F. and Ackers, G. K., Biochem., 29, 6568 (1990).

N-ni-trisodiethanolamine

AG 50W-X8 resin

Wigfield, Y. Y. and Lanouette, M., JAOAC, 68, 1142 (1985).

1. Maniatis, et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Lab (1982).

Metal Ion Removal
Metal Ion Removal
Anions and cations
AG 501-X8
resin

Cations

Anion complexes

pH > 4pH ≤ 4

pH < 2

Chelex 100
resin

AG 50W or
AG MP-50
resin

Chelex 100
resin

AG 1 or
AG MP-1
resin

AG 50W or
AG MP-50
resin

Table 5.3. Guide to Metal Removal
Ion exchange resins are the products of choice for metal concentration andremoval. Many methods of trace metal analysis depend on the prior separation andconcentration of metals from such samples as air, soil, industrial waste water, andbiological extracts. Trace metals can be concentrated directly by adsorption toChelex 100 chelating resin or AG 50W strong cation exchanger, or indirectly
through the adsorption of metal-anion complexes on AG 1 strong anion exchange
resin.

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Mechanism
In ion exchange chromatography, ions are adsorbed by electrostatic forces. Ionsbind to cation and anion exchange resins according to specific selectivities.
Alternatively, Chelex chelating resin contains paired iminodiacetate ions which
act as chelating groups to bind polyvalent metal ions. Chelex has a high preference
for heavy metals over monovalent cations and has a very strong attraction for transition metals. Any metal removed from solution is replaced by an equivalentamount of the resin’s counterion.
Metal cations

Chelex chelating resin has a high selectivity for polyvalent cations and is the preferred method for metal ion removal. Cation adsorption is very low at pH < 2,increases sharply from pH 2-4, and reaches a maximum at a pH > 4. Therefore, for
cation removal, Chelex resin is recommended for use at pH > 4. AG 50W strong
cation exchange resin can also be used for metal concentration, separation, andremoval. However, the selectivity for transition metal cations is very high and elution is difficult.

Metal anion
complexes

The concentration or removal of metal-anion complexes can be achieved throughadsorption on AG 1 anion exchange resin. Alternatively, Chelex 100 resin can beused to bind anion complexes, since it has been found to act as an anion exchanger at pH < 2.2

Metal cation andanion removal

AG 501-X8 mixed bed resin can be used to remove metals and anionic metal
complexes from solutions. Since AG 501-X8 mixed bed resin consists of equivalent amounts of AG 1-X8 strong anion exchange resin and AG 50W-X8 strong
cation exchange resin, it is ideal for removing metals from solutions containing
both anionic metal complexes and metal cations.

Applications
Trace metal removal,
concentration,and metal analysis

Chelex resins’ high affinities for copper, iron, and other heavy metals, as wellas calcium and magnesium ions, make them uniquely suited for removing, concentrating, or analyzing trace metals in solutions, even when large amounts of
sodium and potassium are present. The resins have been used for analysis of trace
metals in natural waters, reagents, biochemicals, physiological fluids, culture
media, soils, and enzyme systems. Trace metals can be concentrated by adsorption
to Chelex chelating resin. The use of Chelex resin to pre-concentrate samples foranalysis has been extensively reviewed.3 Determination of subnanogram levels of
Cd, Co, Cu, Fe, Mn, Ni, Pb, and Zn can be achieved using Chelex 100 resin.4

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Buffer purification

Chelex 100 resin is recommended for removal of heavy metal ion contaminants inbuffers and reagents. It is a very powerful tool in the preparation of high purity
buffers, since it has a very strong attraction for transition metals even in the presence of high salt concentrations.

References
Application

Product

Reference

Boron clean-up

Chelex resin

Gregorie, D., Anal. Chem., 59, 2479 (1987).

C-reactive protein dialysis

Chelex resin

Kinoshita, C. M., et al., Biochem., 28, 9840 (1989).

Calcium removal

Chelex resin

Putnam-Evans, C. L., et al., Biochem., 29, 2488 (1990).

Calcium removal fromsarcoplasmic reticulum vesicle

Chelex resin

Chiesi, M. and Inesi, G., Biochem., 19, 2912 (1980).

Calcium removal fromwhole blood (batch)

Chelex resin

Raymond, F. A. and Weinshilboum, R. M., Clin. Chim. Acta,
58, 185 (1975).

Calmodulin purification

Chelex resin

Vorherr, T., et al., Biochem., 29, 355 (1990).

Chelex competition assay

Chelex resin

Hutnik, C. M. L., et al., Biochem., 29, 7318 (1990).

Disodium salt removal

Chelex resin

Ray, Jr., W. J., et al., Biochem., 29, 2770 (1990).

Divalent cation removal

Chelex resin

Stewart, J. M. McD., et al., Biochem., 28, 4695 (1989).

Divalent cation removalfrom stock solutions

Chelex resin

Devlin, C. C. and Grisham, C. M., Biochem., 29, 6192
(1990).

Equilibrium dialysis

Chelex resin

Thielens, N. M., et al., Biochem., 29, 3570 (1990).

Heavy metal ion removal

Chelex resin

Anderson, V. E. and Cleland, W. W., Biochem., 29, 10498(1990).

Heavy metal ion removalfrom buffer

Chelex resin

Wood, W. M., et al., Biochem., 24, 3686 (1985).

Iron removal from buffer

Chelex resin

Bomford, A., et al., Biochem., 24, 3472 (1985).

Lithium chloride purification

Chelex resin

Ray, Jr., W. J., et al., Biochem., 29, 2770 (1990).

Metal analysis Cu, Cd, Mn,
Zn, Pb from river water

Chelex resin

Liu, Y. and Ingle, Jr., J. D., Anal. Chem., 61, 525 (1989).

Metal analysis Cu, Cd, Znfrom natural waters

Chelex resin

Liu, Y. and Ingle, Jr., J. D., Anal. Chem., 61, 520 (1989).

Metal removal

Chelex resin

Beyer, W. F., et al., Biochem., 28, 4403 (1989); Graf, E., J.
Agric. Food Chem., 31, 851 (1983); Sadhu, A. S. and
Magnuson, J. A., Biochem., 28, 3197 (1989).

Pb removal

Chelex resin

Trost, J. T. and Blankenship, R. E., Biochem., 28, 9898
(1989).

Metal removal from ATP

Chelex resin

Sontheimer, G. M., et al., Biochem., 26, 2701 (1987).

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Application

Product

Reference

Metal removal from ATP
sodium salt

Chelex resin

Avena, R. M. and Bowen, W. J., J. Biol. Chem., 246, 2265
(1971).

Metal removal from buffer

Chelex resin

Chung, H. K. and Ingle, Jr., J. D., Anal. Chem., 62, 2547
(1990); Laue, T. M., et al., Biochem., 28, 4762 (1989).

Metal removal from buffer
biological samples

Chelex resin

Knapp, G., et al., J. Anal. Atomic Spectrometry, 2, 611 and(1987).

Metal removal fromenzyme preparations

Chelex resin

Barker, R., et al., Biochem. J., 177, 289 (1979); Dent, A. J.,
et al., Biochem., 29, 7822 (1990); Dunn, M. F., et al.,Biochem., 19, 718 (1980).

Metal removal from
S100b and melittin

Chelex resin

Baudier, J., et al., Biochem., 26, 2886 (1987).

Oligonucleotide purification

Chelex resin

Kochoyan, M., et al., Biochem., 29, 4799 (1990).

Paramagnetic impurities
removed from oligonucleotides

Chelex resin

Kochoyan, M., et al., Biochem., 29, 4799 (1990).

PCR signal enhancement

Chelex resin

Singer-Sam, J., et. al. Amplifications (Perkin Elmer Cetus)
Issue 3 (1989).

Potassium chloride purification

Chelex resin

Burbaum, J. J. and Knowles, J. R., Biochem., 28, 9306
(1989).

Preirradiation separation
mechanism

Chelex resin

Gokmen, I. G., et al., Anal. Chem., 61, 2757 (1989).

Protease inhibition

Chelex resin

Shogren, R., et al., Biochem., 28, 5525 (1989).

Protein dialysis

Chelex resin

Laue, T. M., et al., Biochem., 28, 4762 (1989).

Protein solution clean-up

Chelex resin

Kaarsholm, N. C., et al., Biochem., 28, 4427 (1989).

Reagent purification for NMR

Chelex resin

Brito, R. M. M., et al., Biochem., 30, 1461 (1991).

Sample preparation

Chelex resin

Schroeder, S. A., et al., Biochem., 28, 8292 (1989).

Sea water analysis

Chelex resin

Pai, S-C., et al., Anal. Chem., 62, 774 (1990).

Selenium determination in fat
materials and petroleum
products

Chelex resin

Narasaki, H., Anal. Chem., 57, 2481 (1985).

Trace metal extraction
from seawater

Chelex resin

Paulson, A. J., Anal. Chem., 58, 183 (1986).

Trace metal separation

AG 50W-X4 resin

Van der Walt, T. N. and Strelow, F. W. E., Anal. Chem., 55,
212 (1983).

Transition metal separation

Chelex resin

Liu, Y. and Ingle, Jr., J. D., Anal. Chem., 61, 520 (1989); Liu,Y. and Ingle, Jr., J. D., Anal. Chem., 61, 525 (1989).

Uridine 5'-diphosphate
chloroacetol synthesis

Chelex resin

Flentke, G. R. and Frey, P. A., Biochem., 29, 2430 (1990).

Vanadium in seawater

Chelex resin

Dupont, V., et al., Anal. Chem., 63, 520 (1991).

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Application

Product

Reference

Vanadium separation from
biological materials

Chelex resin

Fassett, J. D. and Kingston, H. M., Anal. Chem., 57, 2474
(1985).

Water purification

Chelex resin

Grimshaw, C. E., et al., Biochem., 29, 9936 (1990).

2. El Sweify, F. H., Shabana, R., Abdel-Rahman, N. and Aly, H.F., J. Radioanal. Nucl. Chem., 91 (1), 91 (1985).
3. Riley, J. P. and Skirrow, G., Chemical Oceanography, Vol 3, Academic Press, New York (1975).
4. Kingston, H. M., Barnes, I. L., Brady, T. J. and Rains, T. C., Anal. Chem., 50, 2064 (1978).

Anion and Cation Removal
AG 4-X4
resin
Hydrophobic
sampleAG MP-1
resin

Carbohydrate
sampleAG 2-X8
resin

MW 1,000 - 1,400
AG 1-X4
resin

Other
anionsSamplewith anions

MW < 1,000
AG 1-X8
resin

Mixed anions
and cation
sampleAG 501-X8
resinRadioactive
sampleAG MP-1
resin

Organic acidsampleBio-Rex 5
resin

MW 1,400 - 2,700
AG 1-X2
resin
MW > 2,700
AG MP-1
resin

AG 4-X4
resin
Without
proteins
AG 3-X4
resin

Mineral acidsampleWithproteins

Table 5.4. Guide to Anion Removal

38

AG 11 A8resin

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MW < 1,000
AG 50W-X8
resin
Mixed anions
and cation
sampleAG 501-X8
resin

MW 1,000 - 1,400
AG 50W-X4
resin

Other
cations

Metal cation
sampleSamplewith cations

Chelex 100
resin

Organic basesampleBio-Rex 70
resinRadioactive
sampleAG MP-50
resin

Hydrophobic
sample

MW 1,400 - 2,700
AG 50W-X2
resin
MW > 2,700
AG MP-50 resin
or Bio-Rex 70
resin

pH > 6
Bio-Rex 70
resin
pH 2 - 12
AG MP-50
resin

Table 5.5. Guide to Cation Removal
Ion exchange resins can be used to remove or concentrate molecules in a varietyof samples (Tables 5.4 and 5.5). Applications of these resins range from inorganic
ion removal to concentration of large proteins. In general, for zwitterionic compounds, a cationic molecule will be exchanged onto a cation exchanger if the pH
is at least 1 unit below the pI of the cation, and an anion will be exchanged onto
an anion exchanger if the pH is at least 1 unit above the pI of the anion. Inorganic
ions will exchange for the counterion of the resin if the resin selectivity is higher
than that of the counterion. Ion exchange resins allow concentration of dilute
samples and isolation of compounds of interest from complex materials to beaccomplished easily and inexpensively.

Resin Selection Parameters
The following important parameters should be considered when selecting an ion
exchange resin.

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pH

The operative pH range determines which ion exchange resins can be used. Strong
anion exchangers such as AG 1, AG 2, and Macro-Prep high Q resins exchangeions at any pH from 1–14, as do strong cation exchangers, like AG 50W, AG MP
50, and Macro-Prep high S resins. Conversely, weak cation exchangers, like
Bio-Rex 70 and Macro-Prep CM supports, function as cation exchangers only
when their functional groups are ionized (pH > 5). Weak anion exchange resins
such as AG 3-X4 and AG 4-X4 resins also only work when their functional groups
are protonated (pH < 7). When selecting a resin for a particular sample, check the
pH to confirm that the resin functional groups and the sample are oppositely
charged. A zwitterionic compound will be anionic if the pH is at least 1 unit above
its pI and cationic if the pH is 1 unit below its pI.

Ionic form

The ionic form (counterion) of a resin is the ion presently adsorbed by the resin.
The selectivity of the ion to be exchanged (the sample) must be higher than thatof the counterion of the resin. The relative selectivity of various ions is shown in
Table 5.6. If the selectivity of an ion is not known, success can be assured by
selecting the most reactive form of resin; e.g. AG 1 resin in the OH– form or AG
50W resin in the H+ form.

Porosity

The porosity of a support refers to the total pore volume within the matrix of the
support. The greater the pore volume, the greater the porosity. A very porous support may have either many small pores or few large pores. Porous supports withhigh exclusion limits are recommended for high molecular weight compounds
such as proteins and other biomolecules. Low porosity supports are recommended
for low molecular weight compounds such as inorganic ions and organic acids.
Bio-Rad’s high porosity supports include Macro-Prep, Bio-Gel, and Bio-Rex 70
supports. Less porous resins include AG, Dowex, and Chelex resins.

Particle size

Ion exchange resins are available in several particle size ranges. Large particle size
ranges such as 300-1,180 µm, or 20-50 mesh, are recommended for batch techniques because they are easiest to remove from the sample. For column techniques,particle selection is very important and depends on the column size. In general, the38-75 µm particle size (200-400 mesh) is recommended for columns less than 1 ml,
75-150 µm (100-200 mesh) for columns up to 10 ml, 150-300 µm (50-100 mesh)for columns up to 30 ml, and 300-1,180 µm (20-50 mesh) for larger columns.

Time andtemperature

For column applications, a minimum retention time of 1–2 minutes is necessary
for ion exchange to occur. In batch applications, ion exchange continues for up to
24 hours, but for most sample preparation applications 30-60 minutes is sufficient. Ion exchange applications can be performed between 4–85 °C.

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Table 5.6. Relative Selectivity of Various Counterions on AG Resins
AG 50W-X8 resin

AG 1-X8 resin

Counterion

Relative
Selectivity

Counterion

Relative
Selectivity

H+

1.0

OH-

1.0

Li+

0.85

F-

1.6

Na+

1.5

Propionate

2.6

NH4+

1.95

Acetate

3.2

Mn+2

2.35

Formate

4.6

K+

2.5

HPO4-

5.0

Mg+2

2.5

IO3-

5.5

Fe+2

2.55

HCO3-

6.0

Rb+

2.6

Cl-

22

Cs+

2.7

NO2-

24

Zn+2

2.7

BrO3

27

Co+2

2.8

HSO3-

27

Cu+2

2.9

CN-

28

Cd+2

2.95

Br-

50

Ni+2

3.0

NO3-

65

Ca+2

3.9

ClO3-

74

Sr+2

4.95

HSO4-

85

Cu+

5.3

Phenate

110

Hg+2

7.2

I-

175

Pb+2

7.5

Citrate

220

Ag+

7.6

Salicylate

450

Ba+2

8.7

Benzenesulfonate

500

Applications
Ampholyte removal

Ampholytes are amphoteric molecules used in isoelectric focusing. Since
AG 501-X8 mixed bed resin has both anion and cation exchange capabilities, it isideal for removing carrier ampholytes of either positive or negative net charge.
Additionally, mixed bed resins can quantitatively remove carrier ampholytes fromprotein fractions or separate ampholytes from peptides with a molecular weightgreater than 4,000 daltons.

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Organic acids

Low molecular weight anionic molecules such as organic acids are removed using
AG 1-X8 resin. The resin has an approximate molecular weight exclusion limit of
1,000 daltons and is ideal for removing, concentrating, or separating low molecular weight molecules.

Peptides andamino acids

Since peptides and amino acids are amphoteric compounds, they can be concentrated or separated using either AG 1 strong anion exchange resins or AG 50W
strong cation exchange resins. The pI of the molecule and the operating pH will
determine which ion exchange resin to use. Generally, at pH > pI an anion
exchange resin is used, and at a pH < pI, a cation exchange resin is used. The
lower crosslinked resins have higher exclusion limits and are recommended forhigh molecular weight molecules like peptides. Similarly, the higher crosslinked
resins have lower exclusion limits and are recommended for low molecular weightmolecules like amino acids.

Sugars and sugar
alcohols

Sugars and sugar alcohols are optimally deionized using AG 2-X8 strong anion
exchange resin. The resin is similar to AG 1 resin but is capable of deionizing
sugars without isomerization. A stepwise gradient and borate buffers can be used
to separate the sugars.

References
Application

Product

Reference

Acetylglutamate fromglutamate separation

AG 50W-X8 resin

Alonso, E. and Rubio, V., Anal. Biochem., 146, 252 (1985).

Acylcarnitine purification

AG 1-X8 resin

Shinka, T., et al., Kanazawa Ika Daigaku Zasshi, 13, 238(1988).

Adenine nucleotides in blood

AG MP-1 resin

Hsu, D. S. and Chen, S. S., J. Chromatog., 311, 396 (1984).

Adenosine mannoheptose
isolation

Dowex 1-X4 resin

Kocsis, B. and Kontrohr, T., J. Biol. Chem., 259, 11858(1984).

Adenosyl-L-methionine
separation

AG 50W-X4 resin

Miura, G. A. and Chiang, D. K., Anal. Biochem., 147, 217(1985).

Aldehyde and ketone separation

AG 50W-X2 resin

Rendina, A. R. and Cleland, W. W., Anal. Biochem., 117,
213 (1981).

Aldehyde separation

AG 1-X8 resin

Christofferson, K., Anal. Chim. Acta., 33, 285 (1965);
LaNoue, K., et al., J. Biol. Chem., 245, 102 (1970).

Alpha-ketoisocaproic
acid desalting

AG 11 A8 resin

Barrio, J. R., et al., J. Nucl. Med., 24, 515 (1983).

Aluminum concentration

AG 1-X8 resin

Pesavento, M., et al., Analyst., 114, 623-626, (1989).

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Application

Product

ReferenceAmine concentration

AG 1-X8 resin

Minkler, P. E., et al., J. Chromatog., 336, 271 (1984).

Amine measurement in
CNS samples

AG 1-X4 resin
AG 3-X4 resin
Bio-Rex 70 resin

Smith, J. E., et al., Anal. Biochem., 64, 149 (1975).

Amine separation

AG 50W-X8 resin

Charest, R. and Dunn, A., Anal. Biochem., 136, 421 (1984).

Amino acid concentration

AG 50W-X8 resin

Ford, C. W., J. Sci. Food Agric., 35, 881 (1984); Stabler, S. P.,
et al., Anal. Biochem., 162, 185 (1987).

Aminocyclopropane
carboxylic acid separation

AG 50W-X4 resin

Miura, G. A. and Chiang, D. K., Anal. Biochem., 147, 217(1985).

Ammonia determinationin plasma

AG 50W-X8 resin

Forman, D. T., Clin. Chem., 10, 497 (1964).

Ammonium isocyanate
removal from urea

AG 501-X8 resin

Busse, W. and Carpenter, F. H., Biochem., 15, 1649 (1976);
Edelstein, C., et al., Biochem., 15, 1262 (1976); Spieker, H.
and Polet, H., J. Biol. Chem., 251, 987 (1976); Traugh, J. A.
and Porter, G. G., Biochem., 15, 610 (1976).

Ampholyte Removal

AG 501-X8 resin

Bakker, J.A., et al., J. Chromatog., 209, 273, (1981);
Baumann, G. and Crambach, A., Anal. Biochem., 69, 649(1975); Brown,W.D., and Green, S., Anal. Biochem., 34, 593(1975).

Anion removal from
porphyrin in urine

AG 1-X8 resin

Torben, K. and Penderson, J. S., Scan. J. Clin. Lab. Invest.,
38, 279 (1978).

ATP clean-up

Dowex 1-X8 resin

England, P. J., Anal. Biochem., 93, 272 (1979).

ATP from F-actin solution

AG 1 resin

Chock, S. P., et al., Biochem., 15, 3246 (1976).

ATP purity determination

Dowex 1-X8 resin

Hausman, S. Z., et al., Biochem., 29, 6128 (1990).

ATP removal fromproteoliposomes

AG 1-X8 resin

Woldegiorgis, G. and Shrago, E., J. Biol. Chem., 260, 7585
(1985).

Boron clean-up

AG 50W-X8 resin
Chelex resin

Gregorie, D., Anal. Chem., 59, 2479 (1987).

Bound micrococcal
nuclease removal

AG 50W-X2 resin

Goel, S. B. and Modak, S. P., Nucleic Acids Res., 12, 1391
(1984).

Carbohydrate deionization

AG 2-X8 resin
AG 50W-X8 resin

Cullen, M. P., et al., J. Chromatog., 337, 29 (1985).

Carbohydrate removal

AG 1-X8 resin

Marescau, B., et al., J. Chromatog., 377, 334 (1986).

Carboxylated pepsinogen
purification

AG 1-X8 resin

Rajagopalan, T. G., et al., J. Biol. Chem., 241, 4940 (1966).

Carnitine clean-up

AG 1-X8 resin

Minkler, P. E., et al., J. Chromatog., 336, 271 (1984).

Cation removal from sulfate

AG 50W-X8 resin

Hoffer, E. M., et al., Atmospheric Environment, 13, 303(1979).

Chloride uptake of liposomes
determination

Dowex 1-X8 resin

Verkman, A. S., et al., Biochem., 28, 4240 (1989).

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Application

Product

Reference

Creatine kinase isoenzyme
separation (batch)

AG MP-1 resin

Morin, L. G., Clin. Chem., 22, 92 (1976).

Crystalline uric acid preparation

AG 2-X8 resin

Johnson, L. A. and Emmerson, B. T., Clin. Chim. Acta., 41,389 (1972).

Cyclic AMP from cGMP
phosphodiesterase separation

AG MP-1 resin

Hsu, D. S. and Chen, S. S., J. Chromatog., 245, 369 (1982).

Cyclic AMP from cGMP
separation

AG 1-X8 resin

Fallon, A. M. and Wyatt, G. R., Anal. Biochem., 63, 614(1975); Kuehl, Jr., F.A., et al., Proc. Natl. Acad. Sci. USA, 71,
1866 separation (1974).

Cyclic AMP purification

Dowex 50W-X8
resin

Nemecek, G. M., et al., J. Biol. Chem., 254, 598 (1979).

Cyclic nucleotide extraction

AG 1-X8 resin

Schwartzel, Jr., E. H., et al., Anal. Biochem., 78, 395 (1977)

AG 50W-X8 resin

Kuo, W., et al., J. Biol. Chem., 248, 2705 (1973); Schwartz,
D. P., et al., J. Biol. Chem., 248, 2699 (1973).

Cysteinyldopamine fromdicysteinyldopamine

AG 50W-X2 resin

Ito, S. and Fujita, K., J. Chromatog., 375, 134 (1986).

Desformylgramicidin
purification

AG MP-50 resin

Rottenberg, H. and Koeppe,II, R. E., Biochem., 28, 4361
(1989).

Diaminopimelate fromlysine separation

AG 50W-X8 resin

Kelland, J. G., et al., Biochem., 24, 3263 (1985).

Diethyl acetal purification

AG 50W-X8 resin

Cho, Y. K., et al., Biochem., 27, 3320 (1988).

DNA A protein separationfrom E. coli

Bio-Rex 70 resin

Sekimizu, K., et al., J. Biol. Chem., 263, 7136 (1988).

Dopamine hydrochloride
concentration

AG 50W-X12 resin

Miller, S. M. and Klinman, J. P., Biochem., 24, 2114 (1985).

Ethidium bromide removalfrom plasmids

AG 50W-X8 resin

Rodrigues, R. L. and Tait, R. C., Recombinant Techniques—
An Introduction, Addison-Wesley (1983).

Fatty acid removal from lipids

AG 1-X8 resin

Goodridge, A. G., J. Biol. Chem., 248, 4318 (1973).

Free inositol removal frombound inositol

Dowex 1-X8 resin

Grier III, C. E. and Mastro, A. M., J. Immunol., 141, 2585
(1988).

GABA aminotransferase assay

AG 50W-X8 resin

Silverman, R. B. and George, C., Biochem., 27, 3285 (1988).

Gentamicin purificationfrom serum

Bio-Rex 70 resin

Habbal, Z. M., Clin. Chim. Acta, 95, 301 (1979).

Glucose from gluconic acidseparation

AG 1-X8 resin

Marshall, L. M. and Appiah, A., J. Chromatog., 73, 257(1972).

Glucose, sucrose, andfructose extraction

AG 4 resin:
AG 50W resin

Salem, et al., J. Chromatog. Sci., 28, 250-253 (1990).

Glutamic acid purification

Dowex 50 resin

MacKenzie, S. L. and Tenaschuk, J., J. Chromatog., 322, 228(1985).

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Application

Product

Reference

Glycine purification

AG 50W-X8 resin

Arkowitz, R. A. and Abeles, R. H., Biochem., 28, 4639
(1989).

Glycopeptide andoligosaccharide purification

AG 50W-X2 resin

Nishikawa, Y., et al., J. Biol. Chem., 263, 8270 (1988).

Glyphosate quantitation

AG 1-X8 resin
Dowex 50W-X8 resin

Thompson, D., et al., JAOAC, 72, 355 (1989).

Heptose nucleotide
concentration

Dowex 1-X8 resin

Kocsis, B. and Kontrohr, T., J. Biol. Chem., 259, 11858(1984).

I125 cleanup

AG 50W-X8 resin

Auf’mkolk, M., et al., J. Biol. Chem., 261, 11623 (1986).

IDP purification

Dowex 1 resin

Kawakita, N. and Yamazaki, M., Biochem., 17, 3546 (1978).

Inorganic phosphorous
oxo-anion separation

AG 1-X8 resin

Nakamura, T., et al., J. Chromatog., 161, 421 (1978).

Inositol phosphate
determination

AG 1-X8 resin

McCoy, K. L., et al., J. Immunol., 143, 29 (1989).

Inositol phosphate formation

AG 1-X8 resin

Berridge, M. J., et al., J. Biochem., 212, 473 (1983).

Inositol phosphate isolation

AG 1-X8 resin

Grier III, C. E. and Mastro, A. M., J. Immunol., 141, 2585
(1988); Heathers, G.P., et al., Anal. Biochem., 176, 109(1989); Trenn, G., et al., J. Immunol., 142, 3796 (1989).

Dowex 1-X8 resin

Stanley, J. B., et al., J. Immunol., 142, 3546 (1989).

Inositol phosphate separation

AG 1 resin

Smith, C. D., et al., J. Biol. Chem., 262, 6121, (1987).

Inositol removal

Dowex 1-X8 resin

Trenn, G., et al., J. Immunol., 142, 3796 (1989).

L-tryptophan purification

Dowex 50W-X2
resin

Yoshida, R., et al., J. Immunol., 141, 2819 (1988).

Lipase purification

AG MP-1 resin

Baillargeon, M. W. and McCarthy, S. G., Lipids, 26, 831836.

Malate removal fromproteoliposomes

AG 1-X8 resin

Kaplan, R. S. and Pedersen, P. L., J. Biol. Chem., 260, 10293(1985).

Maleic and fumaricacid separation

AG 50W-X4 resin

Richards, M., J. Chromatog., 115, 259 (1975).

Metal absorption
measurement, zinc

AG 1-X8 resin

Eagles, J., et al., Anal. Chem., 61, 1023 (1989).

Metal concentration

AG 1-X8 resin

Porta, V., et al., Mikrochim. Acta., 3, 247-255 (1989).

AG MP-1 resin;
Bio-Beads SM-2 beads

Abollino, O., et al. Anal. Chem., 62, 21-16, (1990).

Methylmalonic acid purification

AG 3-X4 resin

Giorgia, A. J. and Plaut, G. W. E., J. Lab. and Clin. Med., 66,
667 (1985).

Monosaccharide
clean-up-cation removal

AG 50W-X8 resin

Ochiai, M., J. Chromatog., 194, 224 (1980).

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Application

Product

ReferenceN-acetyl-L-[35S]Met
purification

AG 50W resin

Martin, D. J. and Rubenstein, P. A., J. Biol. Chem., 262, 6350
(1987).

Neutral metabolite isolation

AG 1-X8 resin;
AG 50W-X8 resin

Terry, R. C. and Simon, M., J. Chromatog., 232, 261 (1982).

Niacin concentration

AG 1-X8 resin

Tyler, T. A. and Shrago, R. R., J. Liq. Chromatog., 3, 269(1980).

Nitrite determination in meat

AG 50W-X12 resin

Kordorouba, V. and Pelletier, M., Mitt. Geb.
Lebensmittelunters. Hyg., 79, 90 (1988).

Nucleic acid base separationfrom photoproducts

Dowex 1-X8 resin

Salter, L. F. and Thomas, R. C., Biomed. Chromatog., 3, 32(1989).

Nucleic acid separation

AG 1-X8 resin

Heldt, H. W., et al., Anal. Biochem., 101, 278 (1980).

Nucleic acid stripping

AG 50W-X2 resin

Chandrasekaran, E. V., et al., Prep. Biochem., 5, 281 (1975).

Nucleoside mono-, di-,and triphosphate separation

AG 50W-X4 resin

Leigh, C. P. H. and Cashion, P. J., J. Chromatog., 192, 490(1980).

Nucleoside purification

AG 1-X2 resin

Higashi, K., et al., Biochim. Biophys. Acta., 262, 320 (1972);
Tsiftsoglov, A. S. and Georgatsos, J. G., Biochim. Biophys.
Acta., 262, 239 (1972);

Nucleoside separation

AG 1-X2 resin

Asteriadis, G. T., et al., Anal. Biochem., 70, 64 (1976); Kuo,
K. C., et al., J. Chromatog., 378, 361 (1986).

AG 1-X4 resin

Asteriadis, G. T., et al., Anal. Biochem., 70, 64 (1976).

Nucleotide cleanup

AG 50W-X4 resin

DeCamp, D. L., et al., Biochem., 27, 7651 (1988).

Nucleotide separationfrom nucleoside

AG 1-X8 resin

Deutscher, M., J. Biol. Chem., 247, 469 (1972).

Oligogalacturonic acids

AG MP-1 resin

Doner, L.W., et al., J. Chromatog., 449, 229 (1988).

Oligonucleotide separation

AG 1-X2 resin;
AG 1-X4 resin

Asteriadis, G. T., et al., Anal. Biochem., 70, 64 (1976).

Oligouronide fractionation

AG 1-X8 resin

Dave, B. A., et al., J. Chromatog., 116, 395 (1976).

Organic acid concentration

AG 1-X8 resin

Chen, P. M., et al., J. Amer. Soc. Hort. Sci., 107, 807 (1982).

Oxo-L-proline separationfrom proline

AG 50W-X8 resin

Seddon, A. P. and Meister, A., J. Biol. Chem., 261, 11538(1986).

Peptide cleanup

Dowex 1-X2 resin:
Dowex 50W-X2 resin

Schiffmann, E., et al., J. Immunol., 114, 1831 (1975).

Perindoprilate separation

AG 1-X2 resin

van den Berg, H., et al., J. Pharm. Biomed. Anal., 9, 517-524,(1991).

Phosphate mixtures indetergents

AG 1-X8 resin

Lundgren, D. P. and Loeb, N. P., Anal. Chem., 33, 366(1961).

Phosphatidylinositol hydrolysis

AG 1-X8 resin

Wilson, D. B., et al., J. Biol. Chem., 259, 11718 (1984).

Phosphoinositide metabolism

AG 1-X8 resin

Oron, Y., et al., Nature, 313, 141 (1985).

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Application

Product

Reference

Phytate concentration

AG 1-X8 resin

Ellis, R. and Morris, E. R., Cereal Chem., 63, 58 (1986).

Phytic acid concentration

AG 1-X4 resin

Matsunaga, A, et al., Shokuhin Eiseigaku Zasshi, 29, 408(1988).

Phytic acid determination

AG 1-X8 resin

Graf, E. and Dintzis, F. R., J. Agric. Food. Chem., 30, 1094
(1982).

Pi recovery from glucose-6
phosphate

AG 1-X4 resin

Stroop, S. D. and Boyer, P. D., Biochem., 24, 2204 (1985).

Picric acid from proteinhydrolysates

AG 1 resin

Smith, J. E., et al., Anal. Biochem., 64, 149 (1975).

Picric acid removal

AG 1-X8 resin

Rajagopalan, T. G., et al., J. Biol. Chem., 241, 4940 (1966).

Proline concentration

Bio-Rex 70 resin

Larson, A. A. and Dalo, N. L., J. Chromatog., 375, 37 (1986).

Propidium iodide removal

AG 50W-X8 resin

Rodrigues, R. L. and Tait, R. C., Recombinant Techniques—
An Introduction, Addison-Wesley (1983).

Retinoic acid concentration

AG 1-X2 resin

Tamarin, A., et al., J. Embryol. Exp. Morph., 84, 1056 (1984).

Serum thyroxine determination

AG 2-X8 resin

Murphy, B. V. and Jachan, C., J. Lab. and Clin. Med., 66, 161(1965).

Sugar and acid isolationfrom apples

AG 1-X8 resin;
AG 50W-X8 resin

Lee, H. S. and Wrolstad, R. E., JAOAC, 71, 795 (1988).

Sulfamethazine concentration

AG MP-1 resin

Schwartz, D. P., JAOAC, 68, 214 (1985).

Sulfated N-acylhexosamine

AG 1-X8 resin

Nowakowski, R. W., Biochem. Int., 22, 419-426, (1990).
separation

Sulfated proteoglycan
purification

AG 1-X8 resin

Babu, P. B. and Sudhakaran, P. R., J. Cell Biochem., 46, 48-53,(1991).

Sulfathiazole concentration

AG MP-1 resin

Schwartz, D. P. and Sherma, J., JAOAC, 69, 72 (1986).

Taurine clean-up

Dowex 2-X8 resin
Dowex 50W-X8 resin

Stephan, Z. F., et al., J. Biol. Chem., 262, 6069 (1987).

Taurine purification

AG 1-X8 resin
AG 50W-X8 resin

Porter, D. W., et al., J. Chromatog., 454, 311 (1988).

Thyroid hormone removalfrom serum

AG 1-X8 resin

Stanley, F., et al., J. Biol. Chem., 261, 9400 (1966).

Tricarboxylic acid separation

AG 1-X8 resin

Bengtsson, L. and Samuelson, O., Anal. Chim. Acta, 44, 217(1969).

Triiodide removal

AG MP-1 resin

Ikarashi, Y., et al., J. Chromatog., 322, 191 (1985).

Trimethyllysine separationfrom trimethylornithine

AG 1-X8 resin;
AG 50W-X8 resin

Lehman, L. J., et al., Anal. Biochem., 162, 137 (1987).

Tryptamine concentration

Bio-Rex 70 resin

Larson, A. A. and Dalo, N. L., J. Chromatog., 375, 37 (1986).

Urine sample preparation

Bio-Rex 70 resin

Chan, Y. P. and Siu, T. S. S., J. Chromatog., 459, 251 (1988).

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Application

Product

Reference

Urogastrone purification

Bio-Rex 70 resin

Savage, C. R. and Harper, R., Anal. Biochem., 111, 195(1981).

Uronic and biouronic acids

AG 1-X8 resin

Johnson, S. and Samuelson, O., Anal. Chim. Acta., 36, 1(1966).

Vitamin B6 concentration

AG 50W-X8 resin

Tryfiates, G. P. and Sattsangi, S., J. Chromatog., 227, 181(1982).

Weak acid separation

AG 3-X4 resin

Dale, R. A., Clin. Chim. Acta., 41, 141 (1972).

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Organic Compound Removal and Concentration
Sample withorganic
molecules
Separate organic
molecules inorganic solvents
MW 400-14,000

Bio-Beads S-X
beads

Separate organic
molecules fromproteins andnucleic acids
Ionicorganic
molecules

Non-ionic
organic
molecules

AG 50W-X8
resin

Non-polar
compounds

Polar
compounds

AG 3-X4
resin< 40 Å
Bio-Beads
SM-4
adsorbents< 90 Å
Bio-Beads
SM-7
adsorbents

AG 11 A8
resin

40 - 90 Å
Bio-Beads
SM-2
adsorbents

AG MP-50
resin

Table 6.1. Guide to Organic Compound Removal and Concentration

Introduction
Several methods exist for removing or concentrating organic compounds. Either
the organic compound can be bound and the other compounds of interest can beeluted, or the compounds of interest can be bound and the organic compound canbe eluted. In many cases, as in many environmental applications, the organic compound is the compound of interest.

49

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Mechanisms
Hydrophobic Interaction
Bio-Beads SM adsorbents use hydrophobic interaction chromatography to separate
molecules. The adsorbents are neutral, macroporous polymeric beads with highsurface areas for adsorbing organic molecules from aqueous solutions. Bio-Beads
SM-2 and SM-4 adsorbents are used for the adsorption of nonpolar substances.
Alternatively, Bio-Beads SM-7 adsorbents can be used for adsorbing polar substances from nonpolar solvents or for adsorbing nonpolar substances from polar
solvents. Bio-Beads SM adsorbents have been used to remove detergents, emulsifiers, and wetting agents, as well as to separate water soluble steroids, phenols,
drugs, and pesticides.

Gel Permeation
Bio-Beads S-X beads are ideal for gel permeation separations of lipophilic polymers
and other solutes using organic eluants. The beads have exclusion limits from 400
to 14,000 daltons and are particularly useful for separating low molecular weightorganic polymers and other hydrophobic substances. Many different compounds
have been separated on Bio-Beads S-X beads, including pesticides, rodenticides,
polycyclic aromatic compounds, lipids, alkalines, fatty acids, polystyrenes, and avariety of hydrocarbons.

Ion Exchange
Several ion exchange resins can be used to concentrate or separate ionic organic
molecules. For example, AG 11 A8 ion retardation resin has the ability to adsorb
mineral acids while allowing salts and organic molecules to pass through thecolumn. The strong acid adsorption properties of AG 11 A8 resin make it ideal forthe removal of hydrochloric acid from protein hydrolysates.

Applications
Biological Applications
Bio-Beads SM adsorbents are useful for removing polar and nonpolar compounds
from solutions of proteins and nucleic acids, since the macroporous matrix of theadsorbent does not interact with the large biomolecules. Two common biological50

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Page 51

applications are detergent removal with Bio-Beads SM-2 beads and ethidium bromide removal from nucleic acid preparations with Bio-Beads SM beads.
Additionally, Bio-Beads SM-2 beads can be used to remove excess rhodamine andfluorescein from their respective conjugated antibody solutions.

Environmental Applications
Pesticides, PCB,
and PAH

Bio-Beads S-X2 beads are very effective for cleanup of pesticides and polychlorinated biphenyl (PCB) residues in fish extracts.1 Bio-Beads S-X3 beads have beenused for specific separation of chlorinated pesticides from animal fat.2 Clean-up oforganochlorine compounds in dairy products was achieved using Bio-Beads S-X3
resin with a cyclohexane-ethanolacetate (EtOAc) eluant.3 Recoveries were approximately 99% for most of the compounds studied. A rapid, easily automatedmethod for the detection of polycyclic aromatic hydrocarbons (PAHs) in shellfish
such as American lobster and blue mussel uses the gel permeation chromatography
technique on Bio-Beads S-X beads.4 The Bio-Beads procedure is ideal as a screening method in the range of 25–18,000 ng PAH per gram of tissue.

References
Application

Product

Reference

Actinides extraction

Bio-Beads SM-2
beads

Green, L. W., et al., Anal. Chem., 55, 2394 (1983).

Aflatoxin clean-up

Bio-Beads S-X3
beads

Hetmanski, M. T. and Scudamore, K. A., Food Addit.
Contam., 6, 35 (1989).

Aminobenzylphosphonicacid purification

Bio-Beads SM-2
beads

Landt, M., et al., Biochem., 17, 915 (1978).

Anabolic steroids in tissue

Bio-Beads SM-2
beads

Verbeke, R., J. Chromatog., 177, 69 (1979).

Bile acid adsorption

Bio-Beads SM-2
beads

Okuyama, S., et al., Bull. Chem. Soc. Japan, 52, 124(1979); Schwarz, H. P., et al., Clin. Chim. Acta, 50, 197(1974).

Chloramphenicol concentration

AG 50W-X8 resin

Schwartz, D. P. and McDonough, F. E., JAOAC, 67, 563(1984).

Chlorinated hydrocarbons
in water

Bio-Beads SM-2
and SM-4 beads

Picer, N. and Picer, M., J. Chromatog., 193, 357 (1980).

Chlorinated pesticide adsorption

Bio-Beads SM-2
beads

McNeil, E. E. and Otsen, R., J. Chromatog., 132, 277 (1977).

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Page 52

Application

Product

Reference

Chlorophenol adsorption

Bio-Beads SM-2
beads

Grieser, M. D. and Pietrzyk, D. J., Anal. Chem., 45, 1348
(1973).

Cigarette smoke oxygenatedcomponent analysis

Bio-Beads S-X12
beads

Chamberlain, W. J., et al., Anal. Chim. Acta, 111, 235(1979); Snook, M. E., et al., Anal. Chem., 47, 1155 (1975).

Cobalamin adsorption

Bio-Beads SM-2
beads

Fenton, W. A. and Rosenberg, L. E., Anal. Biochem., 90, 119(1978).

Environmental contaminant
analysis

Bio-Beads S-X3
beads

Stalling, D. L., et al., ASTM Special Publication 686, 302(1979).

Fish lipid extract analysis

Bio-Beads S-X3
beads

Burns, B. G., et al., JAOAC, 64, 282 (1981).

Food grade poly (vinyl chloride)
resin fractionation

Bio-Beads S-X3
beads

Gilbert, J., et al., J. Chromatog., 320, 361 (1985);
Waliszewski, S. M. and Szymczynski, G. A., J. Chromatog.,
321, 480 (1985).

Free rhodamine adsorption

Bio-Beads SM-2
beads

Spack, E. G., et al., Anal. Biochem. 158, 233 (1986).

Glucuronide adsorption

Bio-Beads SM-2
beads

Delaborde, S., et al., JHRC and CC, 10, 71 (1987).

Glucuronide adsorption
from water

Bio-Beads SM-4
beads

White, J. D. and Schwartz, D. P., J. Chromatog., 196, 303(1980).

Halogenated contaminant
determination in tissue

Bio-Beads S-X3
beads

LeBel, G. L. and Williams, D. T., JAOAC, 69, 1095 (1985).

Herbicide adsorption from soil

Bio-Beads SM-4
beads

Young, C. C., Proc. Natl. Sci. Counc., ROC (B), 8, 119(1984).

Humic complex concentration

Bio-Beads SM-2
beads

Hiraide, M., et al., Anal. Chim. Acta., 200, 171 (1987).

Hydrocarbon adsorption
from seawater

Bio-Beads SM-2
beads

Gomez-Bellinchon, J. I., et al., Environ. Sci. Technol., 22, 667(1988).

Leukotriene adsorption

Bio-Beads SM-2,
SM-4, SM-7

Salari, H. and Steffenrud, S., J. Chromatog., 378, 35 (1986).

Mycotoxin adsorption from
cereal

Bio-Beads SM-4
beads

Kamimura, H., et al., JAOAC, 64, 1067 (1981).

Nitrophenols adsorption

Bio-Beads SM-2
beads

Grieser, M. D. and Pietrzyk, D. J., Anal. Chem., 45, 1348
(1973).

Organochlorine concentration

Bio-Beads S-X3
beads

Venant, A., et al., Analysis, 12, 266 (1984).

Organophosphate residue
analysis in crops

Bio-Beads S-X3
beads

Ault, J. A., et al., J. Ag. and Food Chem., 27, 825 (1979).

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Application

Product

Reference

Organophosphorous pesticide
determination in food

Bio-Beads S-X3
beads

Blaha, J. J. and Jackson, P. J., JAOAC, 68, 1095 (1985).

PAH determination in shellfish

Bio-Beads S-X3
beads

Musial, C. and Uthe, J. F., JAOAC, 69, 462 (1986).

PCBs adsorption from seawater

Bio-Beads SM-2
beads

Gomez-Bellinchon, J. I., et al., Environ. Sci. Technol., 22, 667(1988).

Pesticide residue analysis

Bio-Beads S-X3
beads

Johnson, L. D., et al., JAOAC, 59, 174 (1976); Steinwandter,
H., Fresenius Z. Anal. Chem., 313, 536 (1982).

Phenol adsorption

Bio-Beads SM-7
beads

Fritz, J. S. and Willis, R. B., J. Chromatog., 79, 107 (1973).

Phenolic acid adsorption
from soil

Bio-Beads SM-4
beads

Young, C. C., Proc. Natl. Sci. Counc., ROC (B), 8, 26 (1984);
Young, C. C., Soil Biol. Biochem., 16, 377 (1984).

Phosphatidylcholine andadsorption

Bio-Beads SM-7
beads

Salari, H., J. Chromatog., 419, 103 (1987).

Phosphatidylethanolamine
adsorption

Bio-Beads SM-2
beads

Salari, H., J. Chromatog., 419, 103 (1987).

Plant growth hormone
adsorption

Bio-Beads SM-7
beads

Andersson, B. and Andersson, K., J. Chromatog. Sci., 242,
353 (1982).

Plant hormone adsorption

Bio-Beads SM-2
beads

Stafford, A. L., et al., J. Chromatog., 294, 485 (1984).

Plasma clean-up

Bio-Beads SM-2
beads

Tamura, M., et al., Biochem., 26, 2797 (1987).

Porphyrin purification

Bio-Beads S-X4
beads

Friley, B. K., et al., J. Chromatog., 258, 310 (1983).

Proline isolation fromfossil bone

Bio-Beads SM-2
beads

Stafford, T. W., et al., Life Sciences, 31, 931 (1982).

Prostaglandin adsorption
from biological fluids

Bio-Beads SM-2
beads

Leffler, C. W., et al., Prostaglandins, 21, 227 (1981).

Pulmonary reaction to IV
injected polymer beads

Bio-Beads S-X8
beads

Schoen, F., et al., J. Biomed. Materials Res., 20, 709 (1986).

Purine, pyrimidine, andnucleoside adsorption

Bio-Beads SM-4
beads

Mills, G. C., J. Chromatog., 242, 103 (1982).

Rodenticide analysis in
animal tissue

Bio-Beads S-X3
beads

Hunter, K., J. Chromatog., 299, 405 (1984); Hunter, K., J.
Chromatog., 321, 255 (1985).

Sludge clean-up

Bio-Beads S-X3
beads

Haile, C. L. and Lopez-Avila, V., USEPA Project Summary
600/S4-84-001, March, 1984.

Steroid adsorption

Bio-Beads SM-4
beads

Shimada, K., et al., J. Chromatog., 378, 17 (1986).

Steroid binding measurements

Hydroxylapatite

Nemoto, T., et al., Biochem., 29, 1880 (1990).

53

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Page 54

Application

Product

Reference

Sulfathiazole concentration

AG MP-1 resin

Schwartz, D. P. and Sherma, J., JAOAC, 69, 72 (1986).

Triadimefon adsorption
from grapes

Bio-Beads SM-2
beads

Nickless, G. and Spitzer, T., J. Chromatog., 208, 409 (1981).

1.
2.
3.
4.

Stalling, et al., JAOAC, 55, 32 (1972).
Ault and Spurgeon, JAOAC, 76, 2 (1984).
Venant, A., et al., Analysis, 12, 266 (1984).
Musial, C. J. and Uthe, J. F., JAOAC, 69, 462 (1986).

54

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Page 55

Columns and Accessories
SampleSmall volume

Intermediate
volumeLarge volume

Poly-Prep columns

Econo-Pac
columns

Econo-Column
chromatography columns

Accessories

Econo
SystemFlow adaptors
and funnels

Racks

Fittings andvalvesTable 7.1. Accessories for Sample Preparation

Introduction
Bio-Rad offers a complete line of low pressure polypropylene and borosilicate glass
chromatography columns, plus a wide selection of column accessories. Accessories
include flow adaptors, reservoirs, funnels, valves, fittings, tubing, and racks; allessential components of high performance, low pressure chromatography.

Columns
Small Volume
Bio-Spincolumns

Bio-Spin columns are disposable, polypropylene columns for centrifuge and gravity
flow chromatography applications. The Bio-Spin columns hold up to 1.2 ml ofchromatography media and can be used with most 1.5 ml microtubes or 12 x 75 mm
test tubes in centrifuge column applications. The columns have bed supports andare completely autoclavable.

55

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Poly-Prep
columns

Page 56

Poly-Prep chromatography columns provide a number of conveniences for sample
preparation and other small scale chromatography applications. The graduated
0.8 x 4 cm columns, constructed of high density polypropylene, hold a standard
bed volume of 2 ml of chromatographic media and include a 10 ml reservoir. At
the bottom of the column is a snap-off seal which leaves a male luer tip whenremoved. Poly-Prep columns are ideal for routine applications in which a small
amount of chromatographic medium is used and then discarded, or for multiple
sample clean-up applications.

Chromatography columns.

Intermediate Volume
Econo-Pac
columnEcono-Pac chromatography columns offer all the features of Poly-Prep columns
with a 20 ml bed volume and a 10 ml reservoir to accommodate larger samples. In
addition, the columns include an upper frit which protects the gel bed from
disruption and prevents the column packing from drying.

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Page 57

Large Volume
Econo-Column
columns

Econo-Column chromatography columns are borosilicate glass columns for low
pressure (< 15 psi) chromatography applications in aqueous solutions. The
columns have translucent end fittings for visualization of the entire the gel bed.
The column tips have male Luer lock end fittings for easy connection to low
pressure systems. Flow adaptors are available for the 1.0, 1.5, 2.5, and 5.0 cm
ID columns.

Systems and Accessories

Automated Econo System.

Econo System
The Econo System brings a higher level of convenience, quality, and performance
to low pressure chromatography than is possible using an assemblage of equipment. Designed for the purification of milligram quantities of proteins andnucleic acids, the Econo System is a series of high quality instruments andprepacked cartridges that combine to create an integrated low pressure chromatography system which delivers outstanding performance at an economical
price. The system can be completely automated, from sample injection to fraction
collection, allowing for walk-away operation.

57

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Page 58

Flow Adaptors
Flow adaptors improve column performance by reducing sample dilution and byprotecting the gel bed. Econo-Column flow adaptors have a three position cam
and a Derlin housing that snaps on to the column. They are available for 1.0, 1.5,
2.5, and 5.0* cm Econo-Column chromatography columns and are compatible
with jacketed Econo-Column chromatography columns as well as Econo-Pac
columns.

Tubing and Fittings
Bio-Rad carries a complete line of fittings, tubing, and valves for plumbing low
pressure chromatography set-ups. Fittings are constructed of polypropylene andexhibit excellent chemical compatibility. Luer fittings can be quickly and easily
fitted to most flexible tubing. Small, medium, and large barbs are available for
0.8 mm, 1.6 mm, and 3.2 mm ID tubing. Bio-Rad supplies silicone, Tygon, andPharMed tubing for any low pressure application.

Poly Column Rack
The Poly Column Rack provides convenience in using Poly-Prep and Econo-Pac
columns. The rack holds up to 20 Poly-Prep columns or 10 Econo-Pac columns,
and includes a collection trough for large volumes of effluent.
*5.0 cm flow adaptor lacks cam mechanism.

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Product Information
Membrane Filters for Particulate Removal
Catalog
NumberProduct Description

Prep-Disc Membrane Filters
343-0002
343-0001
343-0004
343-0005

0.2 µm Prep-Disc Membrane Filter, 50
0.45 µm Prep-Disc Membrane Filter, 50
1.0 µm Prep-Disc Membrane Filter, 50
5.0 µm Prep-Disc Membrane Filter, 50

Micro Prep-Disc Membrane Filters
343-0011
343-0012
343-0014
343-0015

0.2 µm Micro Prep-Disc Membrane Filter, 50
0.45 µm Micro Prep-Disc Membrane Filter, 50
1.0 µm Micro Prep-Disc Membrane Filter, 50
5.0 µm Micro Prep-Disc Membrane Filter, 50

Ion Exchange Chromatography Resins
Catalog
NumberProduct
Description

Ionic
Form

Mesh
Size

Package
Size

Chloride
Chloride
Chloride
AcetateChloride
Chloride
Chloride
ChlorideHydroxide
Chloride
Chloride
AcetateFormateChloride
AcetateFormateChloride
Chloride
Chloride
Chloride50–100100–200
200–400
200–400
50–100100–200
200–400
20–50
20–50
50–100100–200
100–200
100–200
200–400
200–400
200–400
20–50
100–200
200–400
50–100

500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g

Analytical Grade Resins
140-1231
140-1241140-1251140-1253
140-1331
140-1341
140-1351
140-1421
140-1422140-1431
140-1441
140-1443
140-1444
140-1451
140-1453
140-1454
140-2421
140-2441
140-2451
141-0831

AG 1-X2 Resin
AG 1-X2 Resin
AG 1-X2 Resin
AG 1-X2 Resin
AG 1-X4 Resin
AG 1-X4 Resin
AG 1-X4 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 2-X8 Resin
AG 2-X8 Resin
AG 2-X8 Resin
AG MP-1 Resin

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Ion Exchange Chromatography Resins (continued)
Catalog
NumberProduct
Description

Ionic
Form

Mesh
Size

Package
Size

Chloride
ChlorideFree BaseFree Base
HydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenHydrogenH+ & OHH+ & OHH+ & OHSelf adsorbed
Sodium
Sodium
SodiumIron
Sodium
SodiumChloride
Chloride
Sodium
Sodium
Sodium
Sodium100–200
200–400
100–200
100–200
50–100100–200
200–400
50–100100–200
200–400
20–50
50–100100–200
200–400
100–200
200–400
100–200
20–50
20–50
25–35
50–10050–100100–200
200–400
100–200
20–50
20–50
100–200
200–400
20–50
50–100100–200
200–400

500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
500 g
100 g
500 g
10 kg
500 g
500 g
500 g
500 g
500 g
500 g

Chloride
Chloride
FormateHydrogenHydrogen

100–200
200–400
200–400
100–200
200–400

50
50
50
50
50

Analytical Grade Resins (continued)
141-0841
141-0851
140-4341
140-5341
142-1231
142-1241
142-1251142-1331
142-1341
142-1351
142-1421
142-1431
142-1441
142-1451
142-1641
142-1651
143-0841
142-6424
142-6425
142-7425
142-7834
142-2822
142-2832
142-2842
142-2825745-7000745-7001
140-7841
140-7851
142-5822
142-5832
142-5842
142-5852

AG MP-1 Resin
AG MP-1 Resin
AG 4-X4 Resin
AG 3-X4 Resin
AG 50W-X2 Resin
AG 50W-X2 Resin
AG 50W-X2 Resin
AG 50W-X4 Resin
AG 50W-X4 Resin
AG 50W-X4 Resin
AG 50W-X8 Resin
AG 50W-X8 Resin
AG 50W-X8 Resin
AG 50W-X8 Resin
AG 50W-X12 Resin
AG 50W-X12 Resin
AG MP-50 Resin
AG 501-X8 Resin
AG 501-X8(D) Resin
Bio-Rex MSZ 501(D)AG 11 A8 Resin
Chelex 100 Resin
Chelex 100 Resin
Chelex 100 Resin
Chelex 100 Resin
Chelex 20 Resin
Chelex 20 Resin
Bio-Rex 5 Resin
Bio-Rex 5 Resin
Bio-Rex 70 Resin
Bio-Rex 70 Resin
Bio-Rex 70 Resin
Bio-Rex 70 Resin

Prefilled Poly-Prep Columns
731-6211
731-6212731-6221731-6213
731-6214

AG 1-X8 Resin, 2 ml
AG 1-X8 Resin, 2 ml
AG 1-X8 Resin, 2 ml
AG 50W-X8 Resin, 2 ml
AG 50W-X8 Resin, 2 ml

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Ion Exchange Chromatography Resins (continued)
Catalog
NumberProduct
Description

Ionic
Form

Mesh
Size

Package
Size

Hydroxide
Hydroxide
Hydroxide
Hydroxide
Free Base
HydrogenHydrogenHydrogenHydrogenH+ & OHH+ & OHSelf adsorbed
Sodium
Sodium
Sodium200–400
100–200
100–200
200–400
100–200
100–200
200–400
100–200
200–400
20–50
20–50
50–10050–100
200–400
100–200

100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g

Sodium
Sodium
H+ & OHH+ & OH-

100–200
200–400
20–50
20–50

100 g
100 g
100 g
100 g

Biotechnology Grade Resins
143-1255
143-1345
143-2445
143-2446
143-3341
143-5241
143-5341
143-5441
143-5451
143-7424
143-7425
143-7834
143-5832
143-5852
143-2832

AG 1-X2 Resin
AG 1-X4 Resin
AG 1-X8 Resin
AG 1-X8 Resin
AG 4-X4 Resin
AG 50W-X2 Resin
AG 50W-X4 Resin
AG 50W-X8 Resin
AG 50W-X8 Resin
AG 501-X8 Resin
AG 501-X8(D) Resin
AG 11 A8 Resin
Bio-Rex 70 Resin
Bio-Rex 70 Resin
Chelex 100 Resin

Molecular Biology Grade Resins
143-1441
143-1451
143-6424
143-6425

AG 50W-X8 Resin
AG 50W-X8 Resin
AG 501-X8 Resin
AG 501-X8(D) Resin

Macro-Prep Ion Exchange Supports
156-0040
156-0041
156-0042
156-0043
156-0050
156-0051
156-0052
156-0053
156-0030
156-0031
156-0032
156-0033
156-0060
156-0061
156-0062
156-0063
156-0020
156-0021

Macro-Prep High Q Support
Macro-Prep High Q Support
Macro-Prep High Q Support
Macro-Prep High Q Support
Macro-Prep Q Support
Macro-Prep Q Support
Macro-Prep Q Support
Macro-Prep Q Support
Macro-Prep High S Support
Macro-Prep High S Support
Macro-Prep High S Support
Macro-Prep High S Support
Macro-Prep S Support
Macro-Prep S Support
Macro-Prep S Support
Macro-Prep S Support
Macro-Prep DEAE Support
Macro-Prep DEAE Support

61

100 ml500 ml5 liters
10 liters
100 ml500 ml5 liters
10 liters
100 ml500 ml5 liters
10 liters
100 ml500 ml5 liters
10 liters
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Page 62

Ion Exchange Chromatography Resins (continued)
Catalog
NumberProduct Description

Package
Size

Macro-Prep Ion Exchange Supports (continued)
156-0022
156-0023
156-0070
156-0071
156-0072
156-0073

Macro-Prep DEAE Support
Macro-Prep DEAE Support
Macro-Prep CM Support
Macro-Prep CM Support
Macro-Prep CM Support
Macro-Prep CM Support

5 liters
10 liters
100 ml500 ml5 liters
10 liters

Econo-Pac Cartridges
731-0026
731-0027
731-0028
732-0021
732-0025
732-0023
731-0066
731-0067
731-0068
732-0061
732-0065
732-0063
732-0001
732-0005
732-0003

Econo-Pac High Q Cartridge, 5 ml
Econo-Pac High Q Cartridge, 5 ml
Econo-Pac High Q Cartridge, 1 ml
Econo-Pac Q Cartridge, 5 ml
Econo-Pac Q Cartridge, 5 ml
Econo-Pac Q Cartridge, 1 ml
Econo-Pac High S Cartridge, 5 ml
Econo-Pac High S Cartridge, 5 ml
Econo-Pac High S Cartridge, 1 ml
Econo-Pac S Cartridge, 5 ml
Econo-Pac S Cartridge, 5 ml
Econo-Pac S Cartridge, 1 ml
Econo-Pac CM Cartridge, 5 ml
Econo-Pac CM Cartridge, 5 ml
Econo-Pac CM Cartridge, 1 ml

1
5
5
1
5
5
1
5
5
1
5
5
1
5
5

Bio-Gel A Ion Exchangers
153-0740
153-0840

DEAE Bio-Gel A Agarose
CM Bio-Gel A Agarose

250 ml
250 mlHydrophobic Interaction Chromatography Supports
Catalog
NumberProduct Description

Package
Size

Macro-Prep Supports
156-0080
156-0081
156-0082
156-0083
156-0090
156-0091
156-0092
156-0093

Macro-Prep Methyl HIC Support
Macro-Prep Methyl HIC Support
Macro-Prep Methyl HIC Support
Macro-Prep Methyl HIC Support
Macro-Prep t-Butyl HIC Support
Macro-Prep t-Butyl HIC Support
Macro-Prep t-Butyl HIC Support
Macro-Prep t-Butyl HIC Support

62

100 ml500 ml5 liters
10 liters
100 ml500 ml5 liters
10 liters

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Page 63

Hydrophobic Interaction Chromatography Resins (continued)
Catalog
NumberProduct
Description

Wet Particle
Size (µm)

Package
Size

Econo-Pac Cartridges
732-0051
732-0055
732-0053
731-0056
731-0057
731-0058

Econo-Pac Methyl HIC Cartridge, 5 ml
Econo-Pac Methyl HIC Cartridge, 5 ml
Econo-Pac Methyl HIC Cartridge, 1 ml
Econo-Pac t-Butyl Cartridge, 5 ml
Econo-Pac t-Butyl Cartridge, 5 ml
Econo-Pac t-Butyl Cartridge, 1 ml

1
5
5
1
5
5

Bio-Beads SM Adsorbents
152-3920
152-8920
152-3924
152-3934
152-4020
152-4320
152-4334

Bio-Beads SM-2 Beads
Bio-Beads SM-2 Beads,
biotechnology grade
Bio-Beads SM-2 Beads
Bio-Beads SM-2 Beads
Bio-Beads SM-4 Beads
Bio-Beads SM-7 Beads
Bio-Beads SM-7 Beads

300–1,180
300–1,180

100 g
25 g

63–150
63–150300–1,180
300–1,180
63–150

25 g
100 g
100 g
100 g
100 g

Wet Particle
Size (µm)

Package
Size

45–90
45–90
<45
90–18045–90<4590–18045–90<4590–180
90–180
90–180
45–90
90–180
45–90
90–180
45–90
90–180
45–90

100 g
500 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g
1 kg
100 g
100 g
100 g
100 g
100 g
100 g
100 g
100 g

Gel Filtration Chromatography Supports
Catalog
NumberProduct
Description

Bio-Gel P Polyacrylamide Gels
150-4114
150-4115
150-4118
150-4120
150-4124
150-4128150-4130
150-4134
150-4138
150-0738
150-0739
150-4140
150-4144
150-4150
150-4154
150-4160
150-4164
150-4170
150-4174

Bio-Gel P-2 Gel, fine
Bio-Gel P-2 Gel, fine
Bio-Gel P-2 Gel, extra fine
Bio-Gel P-4 Gel, medium
Bio-Gel P-4 Gel,fineBio-Gel P-4 Gel, extra fine
Bio-Gel P-6 Gel, medium
Bio-Gel P-6 Gel, fine
Bio-Gel P-6 Gel, extra fine
Bio-Gel P-6DG Gel, medium
Bio-Gel P-6DG Gel, medium
Bio-Gel P-10 Gel, medium
Bio-Gel P-10 Gel, fine
Bio-Gel P-30 Gel, medium
Bio-Gel P-30 Gel, fine
Bio-Gel P-60 Gel, medium
Bio-Gel P-60 Gel, fine
Bio-Gel P-100 Gel, medium
Bio-Gel P-100 Gel, fine63

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Page 64

Gel Filtration Chromatography Supports (continued)
Catalog
NumberProduct
Description

Wet Particle
Size (µm)

Package
Size

Econo-Pac Columns and Cartridges
732-0011
732-0015
732-6000732-6002
732-6004
732-6006

Econo-Pac P6 Cartridge, 5 ml
Econo-Pac P6 Cartridge, 5 ml
Bio-Spin 6 Chromatography Column
Bio-Spin 6 Chromatography Column
Bio-Spin 30 Chromatography Column
Bio-Spin 30 Chromatography Column

90-18090-18090-18090-18090-18090-180

1
5
10
25
10
25

732-2010

Econo-Pac 10DG Column, 10 ml

90-18030

150–300
75–150
38–75
150–300
75–150
38–75
150–300
75–150
38–75
150–300
75–150
38–75
150–300
75–150500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml500 ml
6 vials

40–80
40–80
40–80
40–80
40–80

100 g
1 kg
100 g
100 g
100 g

Bio-Gel A Agarose Gels
151-0130151-0140
151-0150
151-0430
151-0440
151-0450
151-0730
151-0740
151-0750
151-1030
151-1040
151-1050
151-1330
151-1340
151-1901

Bio-Gel A-0.5m Gel, coarse
Bio-Gel A-0.5m Gel, medium
Bio-Gel A-0.5m Gel, fine
Bio-Gel A-1.5m Gel, coarse
Bio-Gel A-1.5m Gel, medium
Bio-Gel A-1.5m Gel, fine
Bio-Gel A-5m Gel, coarse
Bio-Gel A-5m Gel, medium
Bio-Gel A-5m Gel, fine
Bio-Gel A-15m Gel, coarse
Bio-Gel A-15m Gel, medium
Bio-Gel A-15m Gel, fine
Bio-Gel A-50m Gel, coarse
Bio-Gel A-50m Gel, medium
Gel Filtration Standard, 18 mg

Bio-Beads S-X Beads
152-2150
152-2151
152-2750
152-3350
152-3650

Bio-Beads S-X1 Beads
Bio-Beads S-X1 Beads
Bio-Beads S-X3 Beads
Bio-Beads S-X8 Beads
Bio-Beads S-X12 Beads

Affinity Chromatography Supports
Catalog
NumberProduct Description

Package
Size

Ready-to-Use Affinity Supports
153-7301
153-7302
153-7307153-7304

Affi-Gel Blue Gel, 50–100 meshAffi-Gel Blue Gel, 100–200 mesh
DEAE Affi-Gel Blue Gel
CM Affi-Gel Blue Gel

64

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Affinity Chromatography Supports (continued)
Catalog
NumberProduct Description

Package
Size

Ready-to-Use Affinity Supports (continued)
153-6153
153-6154
153-6159
153-6160
153-6161
153-6162
153-6166
156-0006
156-0005
153-6165
153-6164
153-6173
156-0010
153-5199
153-6101

Affi-Gel Protein A Agarose
Affi-Gel Protein A Agarose
Affi-Gel Protein A MAPS II Kit
Affi-Gel Protein A MAPS II Buffers
Protein A MAPS II Binding Buffer
Protein A MAPS II Elution Buffer
Affi-Gel Protein A MAPS II Regeneration Buffer
Affi-Prep Protein A Support
Affi-Prep Protein A Support
Affi-Prep Protein A MAPS II Kit
Affi-Prep Protein A MAPS II Buffers
Affi-Gel Heparin Gel
Affi-Prep Polymyxin Support
Affi-Gel 501 Gel
Affi-Gel 601 Gel

5 ml
50 ml

5 liters
5 liters
5 liters
5 ml
25 ml

40 ml
25 ml
10 ml
5g

Activated Affinity Supports
153-6099
153-6046
153-6051
153-6052
153-6098

Affi-Gel 10 Gel
Affi-Gel 10 Gel
Affi-Gel 15 Gel
Affi-Gel 15 Gel
Affi-Gel 10/15 Combination

156-0002
156-0001
156-0003
156-0004
153-6047
153-6060
153-6054
153-6055
156-0015
156-0016
156-0017

Affi-Prep 10 Support
Affi-Prep 10 Support
Affi-Prep 10 Support
Affi-Prep 10 Support
Affi-Gel Hz Hydrazide Gel
Affi-Gel Hz Immunoaffinity Kit
Affi-Gel Hz 10x Coupling Buffer ConcentrateAffi-Gel Oxidizer
Affi-Prep Hydrazide Support
Affi-Prep Hydrazide Support
Affi-Prep Hydrazide Support

25 ml
4 x 25 ml
25 ml
4 x 25 ml
2 x 25 mlof each1 x 25 ml
4 x 25 ml
500 ml1000 ml
25 ml
500 ml
250 mg
5 ml
25 ml
500 mlCarbodiimide Activated Supports
153-2401
153-0840
153-0990

Affi-Gel 102 Gel
CM Bio-Gel A Gel
EDAC

65

50 ml
250 ml5g

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Affinity Chromatography Supports (continued)
Catalog
NumberProduct Description

Package
Size

Econo-Pac Columns and Cartridges
732-2026
732-2027
732-0101
732-0105732-0031
732-0035732-2022732-2020
732-0091
732-0093
732-0071
732-0075

Econo-Pac Serum IgG Purification Column, 10 ml
Econo-Pac Serum IgG Purification Kit
Econo-Pac Blue Cartridge, 5 ml
Econo-Pac Blue Cartridge, 5 ml
Econo-Pac DEAE Blue Cartridge, 5 ml
Econo-Pac DEAE Blue Cartridge, 5 ml
Econo-Pac Protein A Column, 2 ml
Econo-Pac Protein A Kit
Econo-Pac Protein A Cartridge, 5 ml
Econo-Pac Protein A Cartridge, 1 ml
Econo-Pac Heparin Cartridge, 5 ml
Econo-Pac Heparin Cartridge, 5 ml

5
1
5
1
5
5
1
5
1
5

Hydroxyapatite Chromatography Supports
Catalog
NumberProduct Description

Package
Size

Bio-Gel Hydroxyapatite Media
130-0150130-0151
130-0420
130-0421
130-0520

Bio-Gel HT Hydroxyapatite
Bio-Gel HT Hydroxyapatite
Bio-Gel HTP Hydroxyapatite
Bio-Gel HTP Hydroxyapatite
Bio-Gel HTP Hydroxyapatite, DNA grade

250 ml500 ml
100 g
1 kg
100 g

Macro-Prep Hydroxyapatite Supports
157-0020
157-0021
157-0025
157-0040
157-0041
157-0045
157-0080
157-0081
157-0085

Macro-Prep Ceramic Hydroxyapatite, 20 µm
Macro-Prep Ceramic Hydroxyapatite, 20 µm
Macro-Prep Ceramic Hydroxyapatite, 20 µm
Macro-Prep Ceramic Hydroxyapatite, 40 µm
Macro-Prep Ceramic Hydroxyapatite, 40 µm
Macro-Prep Ceramic Hydroxyapatite, 40 µm
Macro-Prep Ceramic Hydroxyapatite, 80 µm
Macro-Prep Ceramic Hydroxyapatite, 80 µm
Macro-Prep Ceramic Hydroxyapatite, 80 µm

100 g
1 kg
5 kg
100 g
1 kg
5 kg
100 g
1 kg
5 kg

Econo-Pac Cartridges
732-0081* Econo-Pac HTP Cartridge, 5 ml
732-0085* Econo-Pac HTP Cartridge, 5 ml
732-0083* Econo-Pac HTP Cartridge, 1 ml
* Contains Macro-Prep ceramic hydroxyapatite, Type 2

66

1
5
5

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DNA Purification Supports
Catalog
NumberPackageSize

Product Description

Prep-A-Gene DNA Purification Reagents732-6012
732-6013
732-6010
732-6011
732-6017
732-6019
732-6021
732-6022732-6023
732-6024732-6026

Prep-A-Gene DNA Purification Matrix
Prep-A-Gene DNA Purification Matrix
Prep-A-Gene DNA Purification Kit
Prep-A-Gene DNA Purification Kit
Prep-A-Gene Plasmid Purification Kit
Prep-A-Gene Plasmid Purification Kit
Prep-A-Gene Plasmid Buffer Kit
Prep-A-Gene Binding Buffer
Prep-A-Gene Plasmid Binding Buffer
Prep-A-Gene Wash Buffer
Prep-A-Gene Elution Buffer

2 ml
12 ml
2 ml
12 ml
2 ml
12 ml
500 ml500 ml
250 ml125 mlInstaGene DNA Purification Matrix732-6030

InstaGene DNA Purification Matrix

20 ml

Chromatography Columns
Catalog
NumberID(cm)

Length
(cm)

Area
(cm2)

Max
Vol (ml)

Package
Size

0.20
0.20
0.20
0.20
0.20
0.20
0.20
0.20
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.39
0.79

1
1
2
2
3
3
4
4
2
2
4
4
6
6
8
8
12
12
20
20
4

5
2
5
2
5
2
5
2
5
2
5
2
5
2
5
2
5
2
5
2
5

Standard Econo-Column Columns
737-0506
737-0507
737-0511
737-0512
737-0516
737-0517
737-0521
737-0522
737-0706
737-0707
737-0711
737-0712
737-0716
737-0717
737-0721
737-0722
737-0731
737-0732
737-0751
737-0752
737-1006

0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.7
1.0

5
5
10
10
15
15
20
20
5
5
10
10
15
15
20
20
30
30
50
50
5

67

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Chromatography Columns (continued)
Catalog
NumberID(cm)

Length
(cm)

Area
(cm2)

Max
Vol (ml)

Package
Size

4
8
8
16
16
24
24
40
40
79
103
9
9
18
18
27
27
35
35
53
53
89
89
124
177
230
301
25
25
49
49
98
98
147
147
246
344
491
638
196

2
5
2
5
2
5
2
5
2
2
2
5
2
5
2
5
2
5
2
5
2
5
2
2
2
2
2
5
2
5
2
5
2
5
2
2
2
2
2
1

Standard Econo-Column Columns (continued)
737-1007
737-1011
737-1012
737-1021
737-1022
737-1031
737-1032
737-1051
737-1052
737-1091
737-1093
737-1506
737-1507
737-1511
737-1512
737-1516
737-1517
737-1521
737-1522
737-1531
737-1532
737-1551
737-1552
737-1576
737-1591
737-1593
737-1598
737-2506
737-2507
737-2511
737-2512
737-2521
737-2522
737-2531
737-2532
737-2551
737-2576
737-2591
737-2593
737-5011

1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0

5
10
10
20
20
30
30
50
50
100
120
5
5
10
10
15
15
20
20
30
30
50
50
75
100
120
170
5
5
10
10
20
20
30
30
50
75
100
120
10

68

0.79
0.79
0.79
0.79
0.79
0.79
0.79
0.79
0.79
0.79
0.79
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
1.77
4.91
4.91
4.91
4.91
4.91
4.91
4.91
4.91
4.91
4.91
4.91
4.91
19.63

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Page 69

Chromatography Columns (continued)
Catalog
NumberID(cm)

Length
(cm)

Area
(cm2)

Max
Vol (ml)

Package
Size

19.63
19.63
19.63
19.63

393
589
982
1374

1
1
1
1

0.37
0.79
0.79
1.77

6
12
25
89

1
1
1
1

Standard Econo-Column Columns (continued)
737-5021
737-5031
737-5051
737-5071

5.0
5.0
5.0
5.0

20
30
50
70

Standard Jacketed Econo-Column Columns
737-6108737-6116
737-6131
737-6151

0.7
1.0
1.0
1.5

15
15
30
50

Opened-Ended Jacketed Econo-Column Chromatography Columns, two flow
adaptors included
737-6201
Catalog
Number

1.0

30

0.79

25

1

Product
Description

ColumnID (cm)Functional
Length (cm)Flow Adaptor
Flow Adaptor
Flow Adaptor
Flow Adaptor
Flow Adaptor
Econo-Pac Flow Adaptor

1.0
1.0
1.5
2.5
5.0
1.5

1 to 71 to 14
1 to 14
1 to 14
1 to 14
1 to 7

Flow Adaptors
738-0014
738-0015
738-0016
738-0017
738-0018
738-0019

Glass Reservoirs
737-9112
737-9113

Econo-Column Reservoir, 500 ml
Econo-Column Reservoir, 1,000 ml

Econo-Column Funnels
731-0003

Econo-Column Funnel, 250 ml, 5

Bio-Spin Columns
732-6008

Bio-Spin Chromatography Columns, empty, 100

Poly-Prep Columns
731-1550
731-1555
731-7005
732-8102731-8232

Poly-Prep Columns, empty, 50
Poly-Prep Column Stack Cap, 50
Poly Column Rack, 20 place, with removable tube rack
2-way Stopcock, 10
Female Luer Plug, 25

69

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Chromatography Columns (continued)
Catalog
NumberID(cm)

Length
(cm)

Area
(cm2)

Max
Vol (ml)

Package
Size

Econo-Pac Columns
732-1010
738-0019
732-8102

Econo-Pac Columns, empty, 50
Econo-Pac Flow Adaptor, 1
2-way Stopcock, 10

Econo Systems and Accessories
Catalog
NumberProduct Description

Econo Systems
731-8101

731-8099

Econo System, complete, includes pump, UV monitor, Model 2110 fraction
collector, system controller, rack, and single-pen chart recorder (Model 1325)
Econo System, complete. Same as 731-8101 except 220/240 V
Automated Econo System, includes pump, UV monitor, Model 2128 fraction
collector, system controller, rack, and dual-pen chart recorder (Model 1327),
buffer selector, organizer, and gradient monitor.
Automated Econo System, complete. Same as 731-8114 except 220/240 V

Catalog
NumberProduct
Description

731-8098
731-8114

ID
(mm)

Wall
(mm)

Length(meters)

Tubing731-8210
731-8211
731-8212731-8214731-8215
731-8207
731-8208
731-8209
732-8204
731-8240
731-8241
732-8242
731-8247
731-8248
731-8249
731-8232

Silicone Tubing
0.8
0.8
10
Silicone Tubing
1.6
0.8
10
Silicone Tubing
3.2
0.8
10
Tygon Tubing
0.8
0.8
10
Tygon Tubing
1.6
0.8
10
PharMed Tubing
0.8
0.8
10
PharMed Tubing
1.6
0.8
10
PharMed Tubing
3.2
0.8
10
Teflon Tubing
1.6
0.8
5
Pump Tubing Kit, 20 precut tubes and 0.8 mm ID silicone, 4 sets of fittings
Pump Tubing Kit, 20 precut tubes and 1.6 mm ID silicone, 4 sets of fittings
Pump Tubing Kit, 20 precut tubes and 3.2 mm ID silicone, 4 sets of fittings
Pump Tubing Kit, 20 precut tubes and 0.8 mm ID PharMed, 4 sets of fittings
Pump Tubing Kit, 20 precut tubes and 1.6 mm ID PharMed, 4 sets of fittings
Pump Tubing Kit, 20 precut tubes and 3.2 mm ID PharMed, 4 sets of fittings
Female Luer Plug, 25

Poly Column Rack
731-7005

Poly Column Rack, 20 place,with removable tube rack70

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Page 71

Literature Available
Chromatographic Supports
Bulletin
NumberProduct Information

General Chromatography Applications
1800
1801
1802
1803
1825

Nucleic Acid Analysis Applications
Protein and Peptide Purification Applications
Antibody Purification Applications
Environmental Analysis Applications
Guide to Sample Preparation, 1994

Ion Exchange Chromatography
1075
1224
1441
1747
1752
1840 A-100
1840 A-200
1840 A-300
1840 A-400

DEAE and CM Bio-Gel A Ion Exchange Gels
Chelex 20 Resin for Industrial Heavy Metal Removal
Guide to Ion Exchange (see page 84)
Econo-Pac HTP and Protein A Cartridges
Process Scale Media Guide
Macro-Prep Q Support
Macro-Prep S Support
Macro-Prep CM Support
Macro-Prep DEAE Support

Affinity Chromatography
1066
1085
1092
1099
1107
1298
1424
1429

Affi-Gel 601 Affinity Chromatography Gel
Affi-Gel 10 and 15 Activated Supports
CM Affi-Gel Blue Gel for Protease Free Globulin Fraction From Serum
Immunoaffinity Chromatography
Affi-Gel Blue gel for Enzyme and Blood Protein Purification
Affi-Prep 10 Medium and High Pressure Affinity Matrix
Affi-Gel Hz Immunoaffinity Kit
Affi-Prep Polymyxin Endotoxin Removal Gel

Hydrophobic Interaction Chromatography
1461
1841 B-100

Bio-Beads SM Adsorbent Applications Bibliography
Macro-Prep HIC Support

Hydroxyapatite Chromatography
1115
1842 C-100

HPHT Hydroxyapatite Column
Macro-Prep Ceramic Hydroxyapatite

71

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Chromatography Columns and Accessories
Bulletin
NumberProduct Information

Empty Low Pressure Chromatography Columns
1728

Low Pressure Chromatography Columns and Accessories

Prefilled Low Pressure Columns
1726
1789
1826
1827
1836
1837

Bio-Spin 6 and 30 Spin Chromatography Columns
Econo-Pac Cartridges
Enzyme Purification with the Econo-Pac Q Cartridge
Plasmid Purification with the Econo-Pac Q Cartridge
Antibody Purification with the Econo-Pac Protein A Cartridge and the Econo
SystemEnzyme Purification with the Econo-Pac Q Cartridge

Chromatography Instruments and Accessories
Bulletin
NumberProduct Information

Low Pressure Chromatography Systems
1605
1703
1730

Econo System Technical Specifications
Automated Econo System Components
Purification Systems

Low Pressure Chromatography Detectors
1793

Econo Gradient Monitor

Fraction Collectors
1823
1780

Model 2128 Fraction Collector Recorders
Model 1327 Recorder72

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Page 1

Bio-RadLaboratories

Life ScienceGroup

Bio-Rad Laboratories Main Office, 2000 Alfred Nobel Drive, Hercules, California 94547, Ph. (510) 741-1000, Fx. (510) 741-1060 •Eastern Regional Office, 85A Marcus Dr., Melville, New York 11747, Ph. (516) 756-2575, Fx. (516) 756-2594 •Also in: North Ryde, Australia, Ph. 02-805-5000, Fx. 02-805-1920 • Wien, Austria, Ph. 0222-877 89 01, Fx. 0222-876 56 29 • Nazareth, Belgium,
Ph. 091-85 55 11, Fx. 091-85 65 54 • Mississauga, Canada, Ph. (416) 624-0713, Fx. (416) 624-3019 • Beijing, China, Ph. 2563146, Fx. 2564308 •Paris, France, Ph. 01-49 60 68 34, Fx. 01-46 71 24 67 • München, Germany, Ph. 089-318 84 0, Fx. 089-318 84 100 • Milano, Italy, Ph. 02-21609.1,
Fx. 02-21609.399 • Tokyo, Japan, Ph. 03-3534-7515 Fx. 03-3534-8027 • Veenendaal, The Netherlands, Ph. 08385-40666, Fx. 08385-42216 • Auckland,
New Zealand, Ph. 09-443 3099, Fx. 09-443 3097 • Kowloon, Hong Kong, Ph. 7893300, Fx. 7891257 • Upplands Väsby, Sweden, Phone 46 (0) 8 590-73489,
Fx 46 (0) 8 590-71781 • Madrid, Spain, Ph. (91) 661 70 85, Fx. (91) 661 96 98 • Glattbrugg, Switzerland, Ph. 01/810 16 77, Fx. 01/810 19 33 • Hemel
Hempstead, United Kingdom, Ph. 0800 181134, Fx. 0442 259118

Bulletin 1825 US/EG

93-1124 0194

Printed in USA