Ordering Information
Catalog
Number Product Description
Molecular Weight Standards
161-0303 SDS-PAGE Standards, High, 200 µl
161-0304 SDS-PAGE Standards, Low, 200 µl
161-0317 SDS-PAGE Standards, Broad, 200 µl
161-0314 Silver Stain SDS-PAGE Standards, Low, 200 µl
161-0315 Silver Stain SDS-PAGE Standards, High, 200 µl
161-0306 Biotinylated SDS-PAGE Standards, Low, 250 µl
161-0311 Biotinylated SDS-PAGE Standards, High, 250 µl
161-0319 Biotinylated SDS-PAGE Standards, Broad, 250 µl
161-0320 2-D SDS-PAGE Standards
161-0326 Polypeptide SDS-PAGE Standards, 200 µl
Prestained Standards
161-0305 Prestained SDS-PAGE Standards, Low, 500 µl
161-0309 Prestained SDS-PAGE Standards, High, 500 µl
161-0318 Prestained SDS-PAGE Standards, Broad, 500 µl
161-0324 Kaleidoscope Prestained Standards, 500 µl
161-0325 Kaleidoscope Polypeptide Standards, 500 µl
IEF Standards
161-0310 IEF Standards, pI range 4.45-9.6, 250 µl
Silver Stain
SDS-PAGE Standards,
Low Range
Catalog Number
161-0314
Product shipped on dry ice.
Store at -20 °C upon arrival.
Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules CA 94547
4006047 Rev C
SDS-PAGE Molecular Weight
Standards,Low Range
Specifications
Low Range
Range
Contents Rabbit muscle phosphorylase b
Volume 200 µl concentrated solution
Storage -20 °C
Shelf Life 1 year at -20 °C
Applications 400 full size gels
per vial 800 mini gels
Recommended 12.5%*
gel percentages
14,400 to 97,400 daltons
Bovine serum albumin
Hen egg white ovalbumin
Bovine carbonic anhydrase
Soybean trypsin inhibitor
Hen egg white lysozyme
*Note:
The lowest recommended gel percentage for the low
range standards is 10%. At gel percentages of 10%
or less, one or more of the standards may migrate at
the dye front, depending on running conditions and
other factors. As a result, only four or five bands
may be visible after staining. If this should occur, an
increase in gel percentage is recommended.
Silver Stain SDS-PAGE Standards contain approximately
700 µg total protein in 50% glycerol (w/v), 300 mM NaN3,
20 mM Tris, and 4 mM EDTA. The proteins have been
blended to give bands of equal intensity on SDS polyacrylamide gel systems run according to Laemmli1and stained
with Bio-Rad Silver Stain or Silver Stain Plus. Different
results may be obtained when alternative silver staining
chemistries are used.
Reference
1. Laemmli, U. K., Nature, 227, 680 (1970).
2. Hames, B. D. and Rickwood, D., Gel Electrophoresis of Pro-
teins: A Practical Approach, Second Edition, p. 17, Oxford
University Press, New York (1990).
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2
Protocol
100
90
80
70
60
50
40
30
20
10
Daltons
12108642 cm
Dilute standards 1:20 in SDS Reducing Sample Buffer.*
Heat for 5 minutes at 95 °C. Cool and load 10 µl/well for full
length gels (16-20 cm) or 5 µl/well for mini gels. These load
volumes and dilutions have been optimized for development
with Bio-Rad Silver Stain or Silver Stain Plus for approximately 10 minutes. If silver stain development times vary, the
loading volume or dilution of the standards may need to be
adjusted to optimize the band intensity.
* SDS Reducing Sample Buffer (Prepare imme-
diately before use)
ß-mercaptoethanol 25 µl
Stock sample buffer 475 µl
500 µl
Stock Sample Buffer (Store at room temperature)
Distilled water 4.8 ml
0.5M Tris-HCl, pH 6.8 1.2 ml
Glycerol 1.0 ml
10% (w/v) SDS 2.0 ml
0.1% (w/v) bromophenol blue 0.5 ml
9.5 ml
Use of stock sample buffer with insufficient or old ß-mercaptoethanol may result in doublets at the soybean trypsin
inhibitor and ovalbumin bands.
3
Protein Molecular Weights
Molecular
Protein Weight References
Rabbit muscle 97,400 Titani, K., et. al., Proc. Natl. Acad. Sci.
phosphorylase b USA, 74, 11, 4762 (1977).
Bovine serum 66,200 Brown, J. R., Fed. Proc., 34, 591 (1975).
albumin (BSA)
Hen egg white 45,000 Warner, R. C., "Egg Proteins," in: The
ovalbumin Proteins, Vol. IIA, p. 435 (Neurath, H.
and Bailey, K., eds.), Academic Press,
New York (1954).
Bovine carbonic 31,000 Davis, R. P., "Carbonic Anhydrase," in:
anhydrase The Enzymes, Vol. V, p. 545, (Boyer,
P. D. eds.) Academic Press, New York
(1971).
Soybean trypsin 21,500 Wu, Y. V. and Scheraga, H. A., Bio-chem-
inhibitor istry, 1, 698 (1962).
Hen egg white 14,400 Jolles, P., Angew. Chem., Intl. Edit., 8 227
lysozyme (1969).
4
Phosphorylase b
Bovine serum
album
Ovalbumin
Carbonic
anhydrase
Soybean trypsin
inhibitor
Lysozyme
Fig. 1. Silver Stain SDS-PAGE Standards give bands
of even intensities when stained with Bio-Rad's Silver
Stain or Silver Stain Plus. Note that no extraneous
bands are present. Low range standards run on a 12.5%
SDS polyacrylamide gel and stained with Bio-Rad's Silver
Stain Kit.
5
Fig. 2. Curve generated by plotting the molecular
weight of the low range molecular weight standards
run on a 12% SDS polyacrylamide gel vs. the distance migrated from the interface of the stacking and
separating gels in centimeters. An alternative method
is to plot the log10relative mobility (Rf) vs. the gel concentration, %T, (percentage total monomer, i.e. grams
acrylamide plus bis acrylamide/100ml).
distance migrated by protein
Rf=
distance migrated by dye
The curve can be used to determine molecular weights of
unknown proteins.
2
6
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