Bio-Rad IEF and 2-D Standards - Manual (Page 3)

Constituent Proteins
Color pI MW
Phycocyanin Blue 4.45, 232,000
(3 bands) 4.65,
4.75
ß-Lactoglobulin B 5.1 18,400
Bovine carbonic 6.0 31,000
anhydrase
Human carbonic 6.5 28,000
anhydrase
Equine myoglobin Brown 6.8, 17,500
(2 bands) 7.0
Human hemoglobin A Red 7.1 64,500
Human hemoglobin C Red 7.5 64,500
Lentil lectin 7.80, 49,000
(3 bands) 8.00,
8.20
Cytochrome c Red 9.6 12,200
Note: The pI values given here were determined by
direct measurement with a surface pH electrode.
Instructions for Use
IEF Standards can be applied directly to
IEF gels without any prior treatment. Use 5 µl
for gels which are to be stained with Coomassie
blue, and for silver staining, use 0.5 µl or 5 µl of
a 1:10 dilution of IEF Standards, using deion-
ized, destilled water as a diluent.
3 4
5 6
Introduction
Bio-Rad’s IEF Standards, a mixture of nine
natural proteins with isoelectric points ranging
from 4.45 to 9.6, permit dependable and repro-
ducible pI calibration in IEF gels. The mixture
is provided in a stable aqueous solution. No
reconstitution or dilution is required prior to
use. IEF Standards are intended for use in ana-
lytical acrylamide or agarose isoelectric focus-
ing gels. Five of the nine proteins are naturally
colored to provide continuous monitoring of the
focusing process.
Specifications
Contents Approximately 16.5 mg total
protein in 50% glycerol with
0.02% sodium azide
Volume 250 µl
Storage -20 °C *
Shelf life 1 year at -20 °C
Recommended 5 µl for Coomassie blue
sample volume staining (0.5 µl for silver
stain)
Applications 50 (500 for silver stain)
per vial
* IEF Standards will be shipped at room temperature.
They are stable for transportation at this temperature.
Store at -20 °C upon arrival. Do not store at -70 °C.
Note: IEF Standards are not recommended for 2-D
electrophoresis applications. The proteins are
not naturally colored when denatured by
urea and subsequent electrophoresis will produce
different results than those shown in Figure 1.
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