5969-1576.book Page 1 Wednesday, May 16, 2007 11:26 AM

Agilent Human 14
Multiple Affinity
Removal System
Columns for the
Fractionation ofHigh-AbundantProteins from Human
Proteomic Samples
Instructions

5969-1576.book Page 2 Wednesday, May 16, 2007 11:26 AM

General Information
Introduction
The Agilent Human 14 Multiple Affinity Removal
System consists of a Multiple Affinity Removal System Column based on affinity interactions and optimized buffers for sample loading, washing, eluting,and regenerating. It is specifically designed to fractionate 14 high-abundant proteins from human biological fluids such as plasma, serum, and cerebral
spinal fluid (CSF). This technology enables removalof albumin, IgG, antitrypsin, IgA, transferrin, haptoglobin, fibrinogen, alpha2-macroglobulin,
alpha1-acid glycoprotein, IgM, apolipoprotein AI,
apolipoprotein AII, complement C3, and transthyretin in a single step. The targeted high-abundant proteins are simultaneously removed when crude
biological samples are passed through the column.
Selective immunodepletion provides an enrichedpool of low-abundant proteins for downstream proteomics analysis.
Specific removal of 14 high-abundant proteinsdepletes approximately 94% of total protein massfrom human serum. The low-abundant proteins in
the flow-through fractions can be studied. Removalof high-abundant proteins enables improved resolution and dynamic range for one-dimensional gel
electrophoresis (1DGE), two-dimensional gel electrophoresis (2DGE), and liquid chromatography/

Instructions

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mass spectrometry (LC/MS). The collectedflow-through fractions may need to be concentrated
dependent upon the downstream applications.

Instructions

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Human 14 Multiple Affinity Removal
SystemThe Agilent Human 14 Multiple Affinity Removal
System is specially designed to fractionate thehigh-abundant proteins from the low-abundant proteins, allowing improved analysis of both fractions.
(see Figure 1).

H H
L H L
H L LHL H
H LH L H
Multiple
Affinity
RemovalSystemColumn

HH
H H
H
H HH

Apply Crude Human
Serum/Plasma

Human Proteins Depleted:
Albumin, IgG, Antitrypsin,
IgA, Transferrin, Haptoglobin,
Fibrinogen, Alpha2-Macroglobulin,
Alpha1-Acid Glycoprotein, IgM,
Apolipoprotein AI, Apolipoprotein AII,
Complement C3, Transthyretin

L L
L LL LL
HH
H H
H
H HH

Fraction #1
Low-AbundantProteins

Fraction #2
High-AbundantProteins

Figure 1 The Multiple Affinity Removal System

Instructions

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5969-1576.book Page 5 Wednesday, May 16, 2007 11:26 AM

Product Description
Table 1 Human 14 Multiple Affinity Removal
System Column and Starter Reagent Kit
Product no. Product name Product description
5188-6557
5188-6558
5188-6559

5185-5987

5185-5988

5185-5990

5185-5991

5185-5989

Instructions

4.6 × 50 mm, Remove human albumin,
affinity column IgG, antitrypsin, IgA,
4.6 × 100 mm, transferrin, haptoglobin
affinity column and fibrinogen,
10 × 100 mm, alpha2-macroglobulin,
affinity column alpha1-acid glycoprotein,
IgM, apolipoprotein AI,
apolipoprotein AII, complement C3, transthyretin
Buffer A, 1 L Ready-to-use, optimized
buffer for loading, washing, and equilibrating column
Buffer B, 1 L Ready-to-use, optimized
buffer for elution of boundproteins from column
Spin filters,
For sample cleanup
0.22 µm,
before loading column
1 pack of 25
Concentrators, For concentrating flow5 kDa, MWCO, through fractions
1 pack of 25
Human serum Dilute standard for checkalbumining column capacity55969-1576.book Page 6 Wednesday, May 16, 2007 11:26 AM

Product no. Product name Product description
5185-5995

5021-8814

5185-5986

Replacement Replaced when column
end fitting with inlet frit is clogged.
frit for 4.6 mm
ID column(1 each)Replacement Replace when column
end fitting with inlet frit is clogged.
frit for 10 mm ID
column (1 each)Starter Reagent Kit*
Buffer A: 2 × 1 L
Buffer B: 1 L
Spin filters 0.22 µm: 2 packs of 25
Protein concentrators: 1 pack of 25* Under normal use conditions, the kit should last for approximately 200 injections for 4.6 × 50 mm columns, and
100 injections for 4.6 × 100 mm columns.
CAU T ION

Instructions

Do not expose columns to organic solvents(like alcohols, acetonitrile, etc.), strong oxidizers, acids, or reducing agents, and other protein-denaturing agents. Before attaching thecolumn, purge the LC system and run two
method blank injections according to protocol
to ensure all lines and sample loops are freeof organic solvents.

6

5969-1576.book Page 7 Wednesday, May 16, 2007 11:26 AM

N O TE

Instructions

For LC systems shared with other chemical applications, be sure to purge the LC system first withisopropyl alcohol, and then extensively withwater (approximately 1 hour). After purging, proceed with protocol.

75969-1576.book Page 8 Wednesday, May 16, 2007 11:26 AM

Full Protocol for 4.6 x 50 mm Column(Column capacity: 20 µL human serum/plasma*)
1 Set up Buffer A and Buffer B as the only mobile
phases.
2 Purge lines with Buffer A and Buffer B at a flowrate of 1.0 mL/min for 10 min.
3 Set up LC timetable (see Table 1 for details) andrun two method blanks by injecting 100 µL of
Buffer A without a column.
Ensure proper sample loop size in autosamplers.

N O TE* Consult column certificate of analysis to verify your column capacity.
It is the user's responsibility to adjust loading volume tocompensate for the fact that the concentrations of somehigh-abundant proteins can vary widely depending on thesample origin (source). Several plasma proteins such asalpha1-antitrypsin, haptoglobin, fibrinogen, IgG, andalpha1-acid glycoprotein rise several folds in response tostress, infection, inflammation, or tissue necrosis and are
known as acute phase reactants (Henry, J.B. 1996 - Clinical Diagnosis and Management by Laboratory Methods).
Capacity testing was done with pooled normal plasma
from Rockland Immunochemicals (D519-04).

Instructions

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5969-1576.book Page 9 Wednesday, May 16, 2007 11:26 AM

Table 2 LC method for 4.6 x 50 mm column
Solvent A: Buffer A

Pressure limits: 60 bar

Solvent B: Buffer B
LC Timetable
Time

%B

Flow rateMax.
pressure

1

0.00

0.00

0.125

60

2

9.50

0.00

0.125

60

3

9.51

0.00

1.000

60

4

11.50

0.00

1.000

60

5

11.51

100.00

1.000

60

6

16.00

100.00

1.000

60

7

16.01

0.00

1.000

60

8

25.00

0.00

1.000

60

4 Attach column and equilibrate it in Buffer A for
4 min at a flow rate of 1 mL/min at room temperature.
Instructions

95969-1576.book Page 10 Wednesday, May 16, 2007 11:26 AM

5 Dilute human serum/plasma* 4 times (forexample: 20 µL human plasma with 60 µL of
Buffer A†).
6 Remove particulates with a 0.22 µm spin filter;
1 min at 16,000 g.
7 Inject 80 µL of the diluted serum/plasma at aflow rate of 0.125 mL/min (injected volume
should be adjusted to your column capacity).

14.1958 Collect flow-through fraction (like that whichappears between 5.0–7.5 min in Figure 2) andstore collected fractions at -20 ° C if not analyzed
immediately.
mAU
2500
Bound
(high-abundant proteins)

2000

6.165

1500

Flow-through(low-abundant proteins)

19.152

1000
500
0
0

2.5

5

7.5

10

12.5

15

17.5

20

min

Figure 2 Representative chromatogram of
4.6 x 50 mm column.

* Protocol may be applied to other human biological fluids
like CSF.
† Addition of protease inhibitors in Buffer A for sample
dilution helps prevent protein degradation.

Instructions

105969-1576.book Page 11 Wednesday, May 16, 2007 11:26 AM

9 Elute bound proteins from the column withBuffer B (elution buffer) at a flow rate of
1 mL/min for 5.5 min.
10 Regenerate column by equilibrating it withBuffer A for an additional 9.0 min at a flow rateof 1 mL/min.
11 Store column after equilibrating with Buffer A at
2–8 °C (35–46 °F) in a refrigerator. DO NOTFREEZE THE COLUMN.
12 Analyze. Analyze flow-through containing the
low-abundant proteins. For 1D-SDS-PAGE, an aliquot of the flow-through fraction may be useddirectly. For IEF, 2DGE, and MS analysis of theflow-through fraction, it is necessary to buffer
exchange/desalt to an appropriate buffer. The
5KDa MWCO spin concentrators (part number
5185-5991) may be used for buffer exchange andconcentration. Alternatively, for automateddesalting and concentration, the Agilent
mRP-C18 column (part number 5188-5231) may
be used according to published methods(Agilent Technologies, publication
5989-2506EN).

Instructions

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5969-1576.book Page 12 Wednesday, May 16, 2007 11:26 AM

Full Protocol for 4.6 x 100 mm Column(Column capacity: 40 µL human serum/plasma*)
1 Set up Buffer A and Buffer B as the only mobile
phases.
2 Purge LC system with Buffer A and Buffer B at aflow rate of 1.0 mL/min for 10 min without a column.
3 Set up LC timetable (see Table 3 for details) andrun two method blanks by injecting 200 µL of
Buffer A without a column.
Ensure proper sample loop size in autosamplers.

N O TE* Consult column certificate of analysis to verify your column capacity.
It is the user's responsibility to adjust loading volume tocompensate for the fact that the concentrations of somehigh-abundant proteins can vary widely depending on thesample origin (source). Several plasma proteins such asalpha1-antitrypsin, haptoglobin, fibrinogen, IgG, andalpha1-acid glycoprotein rise several folds in response tostress, infection, inflammation, or tissue necrosis and are
known as acute phase reactants (Henry, J.B. 1996 - Clinical Diagnosis and Management by Laboratory Methods).
Capacity testing was done with pooled normal plasma
from Rockland Immunochemicals (D519-04).

Instructions

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5969-1576.book Page 13 Wednesday, May 16, 2007 11:26 AM

Table 3 LC method for 4.6 x 100 mm column
Solvent A: Buffer A

Pressure limits: 60 bar

Solvent B: Buffer B
LC Timetable
Time

%B

Flow rateMax.
pressure

1

0.00

0.00

0.125

60

2

18.00

0.00

0.125

60

3

18.01

0.00

1.000

60

4

20.00

0.00

1.000

60

5

20.01

100.00

1.000

60

6

27.00

100.00

1.000

60

7

27.01

0.00

1.000

60

8

38.00

0.00

1.000

60

4 Attach column and equilibrate it in Buffer A for
4 min at a flow rate of 1 mL/min at room temperature.
Instructions

13

5969-1576.book Page 14 Wednesday, May 16, 2007 11:26 AM

5 Dilute human serum/plasma* 4 times (forexample: 40 µL human plasma with 120 µL of
Buffer A†).
6 Remove particulates with a 0.22 µm spin filter;
1 min at 16,000 g.
7 Inject 160 µL of the diluted serum/plasma at aflow rate of 0.125 mL/min (injected volume
should be adjusted to your column capacity).

24.026

8 Collect flow-through fraction (like that whichappears between 11–15 min in Figure 3) andstore collected fractions at -20 °C if not analyzed
immediately.
mAU

Bound
(high-abundant proteins)

2500
2000
Flow-through(low-abundant proteins)1000

33.19011.071

1500

500
0
5

10

15

20

25

30

35

min

Figure 3 Representative chromatogram of
4.6 x 100 mm column.

* Protocol may be applied to other human biological fluids
like CSF.
† Addition of protease inhibitors in Buffer A for sample
dilution helps prevent protein degradation.

Instructions

14

5969-1576.book Page 15 Wednesday, May 16, 2007 11:26 AM

9 Elute bound proteins from the column withBuffer B (elution buffer) at a flow rate of
1 mL/min for 7.0 min.
10 Regenerate column by equilibrating it withBuffer A for 11.0 min at a flow rate of 1 mL/min.
11 Store column after equilibrating with Buffer A at
2–8 °C (35–46 °F) in a refrigerator. DO NOTFREEZE THE COLUMN.
12 Analyze. Analyze flow-through containing the
low-abundant proteins. For 1D-SDS-PAGE, an aliquot of the flow-through fraction may be useddirectly. For IEF, 2DGE, and MS analysis of theflow-through fraction, it is necessary to buffer
exchange/desalt to an appropriate buffer. The
5KDa MWCO spin concentrators (part number
5185-5991) may be used for buffer exchange andconcentration. Alternatively, for automateddesalting and concentration, the Agilent
mRP-C18 column (part number 5188-5231) may
be used according to published methods(Agilent Technologies, publication
5989-2506EN).

Instructions

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5969-1576.book Page 16 Wednesday, May 16, 2007 11:26 AM

Full Protocol for 10 x 100 mm Column(Column capacity: 250 µL human serum/plasma*)
1 Set up Buffer A and Buffer B as the only mobile
phases.
2 Purge LC system with Buffer A and Buffer B at aflow rate of 1.0 mL/min for 10 min without a column.
3 Set up LC timetable (see Table 2 for details) andrun two method blanks by injecting 200 µL of
Buffer A without a column.
Ensure proper sample loop size in autosamplers.

N O TE* Consult column certificate of analysis to verify your column capacity.
It is the user's responsibility to adjust loading volume tocompensate for the fact that the concentrations of somehigh-abundant proteins can vary widely depending on thesample origin (source). Several plasma proteins such asalpha1-antitrypsin, haptoglobin, fibrinogen, IgG, andalpha1-acid glycoprotein rise several folds in response tostress, infection, inflammation, or tissue necrosis and are
known as acute phase reactants (Henry, J.B. 1996 - Clinical Diagnosis and Management by Laboratory Methods).
Capacity testing was done with pooled normal plasma
from Rockland Immunochemicals (D519-04).

Instructions

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5969-1576.book Page 17 Wednesday, May 16, 2007 11:26 AM

Table 4 LC Method for 10 x 100 mm Column
Solvent A: Buffer A

Pressure limits: 160 bar

Solvent B: Buffer B
LC Timetable
Time

%B

Flow rateMax.
pressure

1

0.00

0.00

0.50

160

2

20.00

0.00

0.50

160

3

20.01

0.00

1.000

160

4

22.50

0.00

1.000

160

5

22.51

100.00

3.000

160

6

30.00

100.00

3.000

160

7

30.01

0.00

3.000

160

8

40.00

0.00

3.000

160

4 Attach column and equilibrate it in Buffer A for
4 min at a flow rate of 1 mL/min at room temperature.
Instructions

17

5969-1576.book Page 18 Wednesday, May 16, 2007 11:26 AM

5 Dilute human serum/plasma* 4 times (forexample: 200 µL human plasma with 600 µL of
Buffer A†).
6 Remove particulates with a 0.22 µm spin filter;
1 min at 16,000 g.
7 Inject 800 µL of the diluted plasma at a flow rateof 0.5 mL/min (injected volume should be
adjusted to your column capacity).

25.6298 Collect flow-through fraction (like that whichappears between 10.5–14.5 min in Figure 4) andstore collected fractions at -20 °C if not analyzed
immediately.

mAU

Bound
(high-abundant proteins)

2500
2000

12.3241500

Flow-through(low-abundant proteins)

34.482

1000
500
0
5

10

15

20

25

30

35

min

Figure 4 Representative chromatogram of
4.6 x 100 mm column.
* Protocol may be applied to other human biological fluids
like CSF.
† Addition of protease inhibitors in Buffer A for sample
dilution helps prevent protein degradation.

Instructions

18

5969-1576.book Page 19 Wednesday, May 16, 2007 11:26 AM

9 Elute bound proteins from the column withBuffer B (elution buffer) at a flow rate of
3 mL/min for 7.5 min.
10 Regenerate column by equilibrating it withBuffer A for 10.0 min at a flow rate of 3 mL/min.
11 Store column after equilibrating with Buffer A at
2–8 °C (35–46 °F) in a refrigerator. DO NOTFREEZE THE COLUMN.
12 Analyze. Analyze flow-through containing the
low-abundant proteins. For 1D-SDS-PAGE, an aliquot of the flow-through fraction may be useddirectly. For IEF, 2DGE, and MS analysis of theflow-through fraction, it is necessary to buffer
exchange/desalt to an appropriate buffer. The
5KDa MWCO spin concentrators (part number
5185-5991) may be used for buffer exchange andconcentration. Alternatively, for automateddesalting and concentration, the Agilent
mRP-C18 column (part number 5188-5231) may
be used according to published methods (Agilent
Technologies, publication 5989-2506EN).

Instructions

195969-1576.book Page 20 Wednesday, May 16, 2007 11:26 AM

Troubleshooting
It is the user's responsibility to adjust loading volume to compensate for the fact that the concentrations of some high-abundant proteins can vary
widely depending on the sample origin (source).
Several plasma proteins such as alpha1-antitrypsin,
haptoglobin, fibrinogen, IgG, and alpha1-acid glycoprotein rise several folds in response to stress,
infection, inflammation, or tissue necrosis and are
known as acute phase reactants (Henry, J.B. 1996 Clinical Diagnosis and Management by Laboratory
Methods).
Review the following information for troubleshooting your experiments.

High backpressure
A clogged inlet frit may result in high backpressure,distorted peak shape, and diminished column lifetime. To prevent these problems, remove particulates from samples with a spin filter before loading.
If clogging occurs, replace plugged inlet frit (Partnumbers: 5185-5995 for 4.6 mm ID columns and5021-8814 for 10 mm ID columns).

No bound fraction peak
Bound proteins can only be removed from the column by eluting with Buffer B. Check LC timetable toensure enough column time exposure to Buffer B,
for complete removal of bound proteins.
Instructions

205969-1576.book Page 21 Wednesday, May 16, 2007 11:26 AM

Abnormal peak height
Approximately 94% of serum/plasma proteins will
be removed as the bound fraction. The peak heightof the bound fraction is expected to be much greaterthan that of the flow-through fraction. If this order isreversed, two possibilities may be checked:
a Column may not have been regenerated wellenough from previous runs, resulting in lost
capacity. To correct this, elute bound proteins
with Buffer B for 3 additional minutes andre-equilibrate the column with Buffer A.
b Check for signs of biological growth in the
Buffer A reservoir. Replace with fresh Buffer A
for optimal column performance.

Breakthrough of high-abundant proteins inflow-through fraction
The majority (95–99%) of targeted proteins should
be depleted from the sample. If the flow-throughfraction contains unusually high levels of the 14
depleted proteins, two possibilites may be checked:
a The column capacity may have been
exceeded. Reduce the sample load per injection.
b Serum/plasma levels of the targeted proteins
may be unusually high.

Instructions

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Column Specifications
Table 5 Column Descriptions
Parameter

Description

Part number 5188-6557

4.6 mm × 50 mm(0.83 mL)

Part number 5188-6558

4.6 mm × 100 mm(1.66 mL)

Column body material

Polyetheretherketone(PEEK)Part Number 5188-6559

10 mm × 100 mm(7.85 mL)

Column body material

PEEK encapsulated instainless steel

End-fitting materials

PEEK with 2 µm frits

Column capacity*

0.83 mL column: 20 µL
human plasma
1.66 mL column: 40 µL
human plasma
7.85 mL column: 250 µL
human plasma

Max. pressure

60 or 160 bar

Instructions

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Table 5 Column Descriptions
Parameter

Description

Operating temperature

18–25 °C

Column packing material

Affinity resin

Immobilized ligands

Affinity ligands to humanalbumin, IgG, antitrypsin, IgA, transferrin, haptoglobin and fibrinogen,alpha2-macroglobulin,
alpha1-acid glycoprotein,
IgM, apolipoprotein AI,
apolipoprotein AII, complement C3, transthyretin

Flow rate range

0.125–3.0 mL/min

Shipping solution

Buffer A with < 0.02%
sodium azide

Shipping temperature

2–8 °C (35–46 °F)

Storage temperature

2–8 °C (35–46 °F)* For exact column capacity, consult your column certificate of analysis.

Instructions

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Safety Issues
When preparing biological samples using any Agilent Multiple Affinity Removal System Column, follow general guidelines for handling biologicalmaterials and wear protective eyewear and gloves.

Recommendations• Sample dilution
It is not recommended to load crude
serum/plasma directly onto the column. Follow
instructions for serum/plasma dilution (fourtimes dilution with Buffer A). Addition of protease inhibitors in Buffer A for sample dilution
helps prevent protein degradation.
• Sample cleanup
Human serum/plasma may contain particulatematerials that can be removed by a quick spinusing a 0.22-µm spin filter.
• Column performance
Always use Buffers A and B as the mobile phases
according to the LC protocols. Do not expose thecolumns to solvents other than Buffers A and B.
• Column storage
After equilibrating Agilent Multiple Affinity
Removal System Columns with Buffer A, always
store them in a refrigerator at 2–8 °C (35–46 °F)
when not in use to minimize loss in column
capacity.
Instructions

24

5969-1576.book Page 25 Wednesday, May 16, 2007 11:26 AM• Lyophylization of flow-through fractions
Buffer exchange to a volatile buffer (for example,
ammonium bicarbonate) is recommended prior tolyophylization due to high salt concentration in
Buffer A.
• Multiple Affinity Removal System
The Agilent Multiple Affinity Removal System is
specially designed to allow for close study of
low-abundant proteins present in flow-throughfractions (see Figure 1).

Do not expose columns to organic solvents(like alcohols, acetonitrile, etc.), strong oxidizers, acids, or reducing agents, and other protein denaturing agents.

CAU T ION

For RESEARCH USE ONLY. This product is
NOT TO BE USED AS AN IN-VITRODIAGNOSTIC.

WARN IN G

Storage: Upon its receipt and when you are notusing it, store the column with the end-caps tightly
sealed in a refrigerator at 2–8 °C (35–46 °F). DO NOTFREEZE THE COLUMN.
For more information or technical assistance, pleasecall toll free: 1-800-227-9770 or visit our Web site at:www.agilent.com/chem/bioreagents.
Instructions

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5969-1576.book Page 26 Wednesday, May 16, 2007 11:26 AM

Related Agilent Products
Other related Agilent products include the following:
5188-6560 Agilent Multiple Affinity Removal System
Spin Cartridge Hu-14, spin cartridge that
depletes 14 high-abundant proteins (albumin, IgG, IgA, transferrin, haptoglobin, antitrypsin, fibrinogen, alpha2-macroglobulin,
alpha1-acid glycoprotein, IgM, apolipoprotein AI, apolipoprotein AII, complement C3,
and transthyretin) from human serum/plasma samples, 8–10 µL serum/plasma
capacity per use
5188-6409 Agilent High Capacity Multiple Affinity
Removal System Column Hu-7HC,
4.6 X 50 mm, LC column that depletes
seven high-abundant proteins (albumin,
IgG, IgA, transferrin, haptoglobin, antitrypsin, and fibrinogen) from human serumor plasma samples, 30–35 µL
serum/plasma capacity per injection
5188-6410 Agilent High Capacity Multiple Affinity
Removal System Column Hu-7HC,
4.6 X 100 mm, LC column that depletes
seven high-abundant proteins (albumin,
IgG, IgA, transferrin, haptoglobin, antitrypsin, and fibrinogen) from human serumor plasma samples, 60–70 µL
serum/plasma capacity per injection

Instructions

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5969-1576.book Page 27 Wednesday, May 16, 2007 11:26 AM

5188-6411 Agilent High Capacity Multiple Affinity
Removal System Column Hu-7HC,
10 X 100 mm, LC column that depletes
seven high-abundant proteins (albumin,
IgG, IgA, transferrin, haptoglobin, antitrypsin, and fibrinogen) from human serumor plasma samples, 250–300 µL
serum/plasma capacity per injection
5188-6408 Agilent High Capacity Multiple Affinity
Removal System Spin Cartridge Hu-7HC,
spin cartridge that depletes sevenhigh-abundant proteins (albumin, IgG, IgA,
transferrin, haptoglobin, antitrypsin, andfibrinogen) from human serum or plasma
samples, 12–14 µL serum/plasma capacity
per use5188-5332 Agilent High Capacity Multiple Affinity
Removal System Column Hu-6HC,
4.6 X 50 mm, LC column that depletes six
high-abundant proteins (albumin, IgG, IgA,
transferrin, haptoglobin, and antitrypsin)from human serum/plasma samples,
30–40 µL serum capacity per injection
5188-5333 Agilent High Capacity Multiple Affinity
Removal System Column Hu-6HC,
4.6 X 100 mm, LC column that depletes six
high-abundant proteins (albumin, IgG, IgA,
transferrin, haptoglobin, and antitrypsin)from human serum/plasma samples,
60–80 µL serum capacity per injection

Instructions

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5969-1576.book Page 28 Wednesday, May 16, 2007 11:26 AM

5188-5336 Agilent High Capacity Multiple Affinity
Removal System Column Hu-6HC,
10 X 100 mm, LC column that depletes six
high-abundant proteins (albumin, IgG, IgA,
transferrin, haptoglobin, and antitrypsin)from human serum/plasma samples,
300–325 µL serum capacity per injection
5188-5341 Agilent High Capacity Multiple Affinity
Removal System Spin Cartridge Hu-6HC,
spin cartridge that depletes six high-abundant proteins (albumin, IgG, IgA, transferrin,
haptoglobin, and antitrypsin) from humanserum/plasma samples, 14–16 µL serum
capacity per use
5185-5984 Multiple Affinity Removal System Column
Hu-6, 4.6 × 50 mm, LC column that
depletes six high-abundant proteins (albumin, IgG, IgA, transferrin, haptoglobin, andantitrypsin) from human serum/plasma
samples, 15–20 µL serum capacity per
injection
5185-5985 Multiple Affinity Removal System Column
Hu-6, 4.6 × 100 mm, LC column that
depletes six high-abundant proteins (albumin, IgG, IgA, transferrin, haptoglobin, andantitrypsin) from human serum/plasma
samples, 30–40 µL serum capacity per
injection

Instructions

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5969-1576.book Page 29 Wednesday, May 16, 2007 11:26 AM

5188-2714 Multiple Affinity Removal System Column
Hu-6, 10 × 100 mm, LC column that
depletes six high-abundant proteins (albumin, IgG, IgA, transferrin, haptoglobin, andantitrypsin) from human serum/plasma
samples, 140–190 µL serum capacity per
injection
5188-5230 Multiple Affinity Removal System Spin
Cartridge Hu-6, spin cartridge that depletes
six high-abundant proteins (albumin, IgG,
IgA, transferrin, haptoglobin, and antitrypsin) from human serum/plasma samples, 7–10 µL serum capacity per use
5188-5217 Multiple Affinity Removal System Column
Ms-3, 4.6 × 50 mm, LC column that
depletes three high-abundant proteins(albumin, IgG, and transferrin) from mouseserum/plasma samples, 37–50 µL serum
capacity per injection
5188-5218 Multiple Affinity Removal System Column
Ms-3, 4.6 × 100 mm, LC column that
depletes three high-abundant proteins(albumin, IgG, and transferrin) from mouseserum/plasma, 75–100 µL serum capacity
per injection
5188-5289 Multiple Affinity Removal System Spin
Cartridge Ms-3, spin cartridge that
depletes three high-abundant proteins(albumin, IgG, and transferrin) from mouseserum/plasma samples, 25–30 µL serum
capacity per use

Instructions

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5185-5986 Multiple Affinity Removal System Reagent
Kit, starter reagent kit containing buffers,
spin filters, and spin concentrators for use
with Multiple Affinity Removal System LC
Columns
5188-5254 Starter Reagent Kit for Spin Cartridges*,
Buffer A: 1 L
Buffer B: 1 L
Spin filters 0.22 µm: 2 packs of 25
Protein concentrators: 1 pack of 25
Luer-Lock adapters: 1 pack of 2
5-mL plastic Luer-Lock syringes: 1 pack of 2
1.5-mL microtubes: 6 packs of 100
Spin cartridge extra caps and plugs, 1 packof 6 each
Teflon Luer-Lock needles, 1 pack of 105188-5231 mRP-C18 High Recovery Protein Fractionation and Desalting Column, seewww.agilent.com/chem/bioreagents formore details* Under normal conditions, the kit should last for approximately 200 spin cartridge uses.

For more information or technical assistance, pleasecall toll free: 1-800-227-9770 or visit our Web site at:www.agilent.com/chem.

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5969-1576.book Page 32 Wednesday, May 16, 2007 11:26 AM©Agilent Technologies, Inc. 2007
Agilent shall not be liable for errors
contained herein or for incidental orconsequential damages in connection
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use of this material.
Information, descriptions, and specifications in this publication are subject
to change without notice.
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May 20075969-1596