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Lonza Rockland, Inc.
www.lonza.com
biotechserv@lonza.com
Tech Service: 800-521-0390
Customer Service: 800-638-8174
Document # 18842-1007-02
Rockland, ME 04841 USASeaPrep® Agarose
An ultralow gelling, soft agarose

Introduction
SeaPrep® Agarose is a specialty “soft agarose” productwith ultralow gelling and melting temperature properties
which make it valuable for many techniques involving
biomolecules. SeaPrep® Agarose is an excellent
medium for hybridoma cloning in soft, sparkling cleargels. (The clear sol may be stored sterile, almost
indefinitely, in a 37ºC incubator.) SeaPrep® Agarosegels below 17ºC and then may be melted at 40ºC-50ºC.
The extended ultralow working temperature range
permits great latitude and convenience for handling ofheat-labile biomolecules.
At concentrations of 0.8%-1.5%, the soft gel texture
makes SeaPrep® Agarose a suitable suspension
medium for cells and facilitates their recovery by gentle
mechanical disruption for subsequent growth in liquid
media. Growth factors can be added to SeaPrep®
Agarose along with other nutrients to produce a“defined” medium. Prepurchase samples of SeaPrep®
Agarose are available for evaluation of compatibility withspecific cell lines.
Analytical Specifications
Gelling temp. (0.8%)
Melting temp. (1.0%)
Moisture:Sulfate:EEO(-mr):Gel strength (2%):

Figure 1. Gelling temperature as a function of the degree ofhydroxyethylation.

Two consequences of the hydroxyethylation of agarose
are a reduction in gel strength and an increase of gel
clarity. The relationship between gel strength andgelling temperature is illustrated in Figure 2.

8ºC-17ºC≤ 50ºC≤ 10%≤ 0.10%≤ 0.05≥ 75g/cm2

Properties
SeaPrep® Agarose is prepared by the controlled
introduction of hydroxyethyl groups to the agarose
molecule (Figure 1). The reduction in gelling
temperature from the natural agarose value ofapproximately 36ºC is a function of the degree ofhydroxyethylation (Figure 1).

Figure 2. Relationship between gelling temperature and gel
strength.

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SeaPrep® Agarose falls near the bottom of the curve in
Figure 2, with a 1% gelling temperature below about20ºC.

Agarose Preparation
In some cases, such as hybridoma cloning when using
SeaPrep® Agarose it is advisable to sample different lotsof agarose for the desirable gel strength qualities.

Gelling temperature, melting temperature, and gel
strength all vary considerably with concentration of
SeaPrep® Agarose in a gel (Figures 3 and 4). In Figure
4, the effect of concentration is particularly pronounced
at concentrations below 1.5%. SeaPrep® Agaroseshould be used above 2% if a usable gel structure isdesired. At higher levels, the remarkable clarity ofSeaPrep® Agarose remains, allowing the investigator toeasily view samples, such as cells, within gels. In
horizontal electrophoretic applications, concentrations
above 2% are recommended.

When preparing agarose for cell culture work, it isalways best to prepare the agarose in water suitable forcell culture and separate from any growth media or
nutrients. Agarose solution and media solutions should
be prepared at 2X concentrations (i.e., if desired final
agarose concentration is 0.6%, prepare a 1.2% agarose
solution), autoclaved separately, and aliquoted into
useable aliquots.
General Instructions Dissolving
1.

7060504030201002.

MeltingGelling

3.
0246

4.

8Concentration (%) Sea Prep Agarose

5.

Figure 3. Relationship between gelling/melting temperature®

and gel concentration of SeaPrep Agarose.

6.

Measure an appropriate volume of water or buffer(room temperature) into a beaker or wide mouthflask that is 2-4 times the volume of the solution
to be heated.
Slowly sprinkle a weighed amount of SeaPrep®
agarose powder into the water or buffer with rapid
stirring such as with a magnetic stir bar.
Weigh the vessel and its contents, then cover itwith vented plastic wrap.
Heat to boiling with continuous rapid stirring.
SeaPrep® agarose sometimes lumps, so gentle
boiling with stirring may be required for 20-30
minutes.
Remove the plastic wrap, weigh the vessel andcontents and correct for evaporation.
Cool the solution. SeaPrep® agarose solutions canbe poured at room temperature. Poured gelsshould be cooled below their gelling temperature(refrigerated) for at least two hours to achieve
maximum gel strength.

Microwave Dissolving
Instructions and timing are based on a single 250 ml
flask containing 100 mL of solution in a 720-watt oven.
Adjustments should be made for different solution
volumes and/or multiple solutions in the microwave
oven.
1.
Disperse agarose as usual and weigh container.
2.
REMOVE MAGNETIC STIR BAR and heat thecovered vessel in the microwave oven at MEDIUM(70%) power setting for 2 minutes.
3.
GENTLY swirl to resuspend any settled powder
and gel pieces.
4.
Reheat at HIGH (100%) for 1-2 minutes until thesolution comes to a boil.
5.
GENTLY swirl to thoroughly mix contents.
6.
Remove plastic wrap and adjust for evaporation.

Figure 4. Relationship between gel strength and gel®
concentration of SeaPrep Agarose.
Cell Biology Applications• Soft agarose cloning of hybridomas1 and humantumor cells2,3.
• General cell culture4; plant protoplast culture5.
• Embedding/encapsulation of cells for electron
microscopy6, neuron activity studies7, transplantation
research in rats8.
• Electrophoresis of cells in a density gradient withreversible gel9.

Microwave Melting of Gels
1.
Loosen cap of container.
2.
Set container in microwave oven and heat at
DEFROST (50%) power setting for 1 minute (Thisis for a 100-mL gel volume; use shorter times forsmaller volumes.)
23.

4.

GENTLY swirl contents to break up gel matrix. If
necessary, reheat at DEFROST briefly until all gel
pieces are melted.
Cool to an appropriate temperature.

Procedure for Autoclaving Agarose
1.
2.
3.
4.
5.
6.

7.

Choose a flask that is 2-4 times the volume of thesolution.
Add water to the flask.
Sprinkle in the pre-measured agarose powder at a2X final agarose concentration.
Cover the flask with aluminum foil.
Place the flask in the autoclave.
Sterilize the agarose by autoclaving for 5 minutes
at 15 lb/in2.
Note: Agarose may lose gel strength when
exposed to longer periods of time in the autoclave.
Once the agarose solution has cooled, aliquot into
useable aliquots and store at 4ºC prior to use.

Order Information:

Catalog No.
50302

For more information contact Technical Service at(800) 521-0390 or visit our website at www.Lonza.com
Related Products:
Lonza Cells and MediaFor Research Use Only. Not for use in diagnostic
procedures.

General Procedure for Using Agarose in Culture
Medium1.
2.
3.
4.

5.
6.

Size
25g

Remelt the agarose by placing in a hot water bathor microwave.
Allow the agarose solution to cool to 37ºC.
Prewarm the 2X media solution to 37ºC.
Mix equal volumes of the sterile 2X agarose
solution with sterile 2X media containing growth
factors and nutrients.
Cast the agarose/media solution into plates orsterile culture tubes.
Allow the agarose solution to gel for 20 minutes if
using as a feeder or overlay or maintain thesolution at 37ºC if using as a liquid culture.

References
Civin, C.I; and Banquerigo, M.L. Journal of
Immunological Methods 1983, 61, 1-8.
2.
Pavlik, E.J.; et al. InVitro 1983, 19,538-550.
3.
Bagasra, O.; et al. Cancer Immunology andImmunotherapy 1985, 20, 55-60.
4.
Liebow, C.; et al. Digestive Diseases andSciences 1983, 28, 940.
5.
Adams, T.L; and Townsend, J.A. Plant Cell
Reports 1983,2,165-168.
6.
Stausbauch, P.;et al. Journal of Electron
Microscopy Techniques 1985,2,261-262.
7.
Mooney, R.; and Waziri, R.; Journal of
Neuroscience Methods 1982,5,249-251.
8.
Howell, S.L.; et al. Journal of Physiology 1982,324, 20P.
9.
Plank, L.D.; et al. presentation at “Electrophoresis84”, Univ. of Arizona, Tucson, AZ, 1984.
1.

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