User guide

Protein Molecular Weights
The molecular weights of every lot of
prestained standards are individually calibrated
against Bio-Rad’s SDS-PAGE Standards. The
lot specific calibrated molecular weights are
included with every vial. Prestained standards
are useful for estimating the molecular weights
of sample proteins, however, for precise molec-
ular weight determination, use Bio-Rad’s SDS-
PAGE, Silver Stain SDS-PAGE, or
Biotinylated SDS-PAGE Standards in addition
to the prestained standards.
Constituent Proteins
(See enclosed insert for lot specific
calibrated molecular weights.)
Protein Source
Myosin Rabbit skeletal muscle
ß-galactosidase E. coli
Bovine serum albumin Bovine plasma
Ovalbumin Chicken egg white
Covalently bound dye alters the molecular weight of the
proteins and produces relatively broad bands. The molecular
weights are calibrated from the center of each protein band.
34
5 6
Myosin
β-galactosidase
Bovine serum
albumin
Ovalbumin
Instructions for Use
Heat the solution to 40 °C for 1 minute
to dissolve any solids which may have pre-
cipitated at -20 °C. To visualize the
prestained standards after blotting, load 10 µl
for full size gels (16-20 cm) and 5 µl for
mini gels. To visualize the standards during
electrophoresis, load 20 µl for full length
gels and 10 µl for mini gels. To see the stan-
dards during the run, it is helpful to hold a
sheet of white paper behind the gel.
Note
The prestaining of the proteins substan-
tially inhibits them from being further stained
with biotin/avidin systems, colloidal gold,
Coomassie R-250, or amido black. The stan-
dards can be silver stained, but silver staining
will result in broad bands because of the large
amount of protein in the sample.
Fig. 1. High Range Prestained SDS-PAGE Standards.
5 µl of the standards were run on a 7.5% SDS poly-
acrylamide gel according to the method of Laemmli.
1
The standards were run on the Mini-PROTEAN
®
II cell
and electrophoretically transferred to nitrocellulose
using the Mini Trans-Blot
®
cell.
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