Bio-Rad IEF and 2-D Standards User Manual

IEF Standards

Instruction Manual

Catalog Number 161-0310

Broad Range pl 4.45-9.6

Product shipped at room temperature.

For research use only.

Store at -20 °C.

Ordering Information

Catalog
Number Product Description

Specialty Standards

161-0310 IEF Standards, pI range 4.45-9.6, 250 µl
161-0320 2-D SDS-PAGE Standards, 500 µl

Prestained Standards

161-0324 Kaleidoscope Prestained Standards, 500 µl
161-0325 Kaleidoscope Polypeptide Standards, 500 µl
161-0305 Prestained SDS-PAGE Standards,

low range, 500 µl

161-0309 Prestained SDS-PAGE Standards,

high range, 500 µl

161-0318 Prestained SDS-PAGE Standards,

broad range, 500 µl

SDS-PAGE Molecular Weight Standards

161-0304 SDS-PAGE Standards, low range, 200 µl
161-0303 SDS-PAGE Standards, high range, 200 µl
161-0317 SDS-PAGE Standards, broad range, 200 µl
161-0326 Polypeptide Standards, 200 µl
161-0314 Silver Stain Standards, low range, 200 µl
161-0315 Silver Stain Standards, high range, 200 µl
161-0306 Biotinylated Standards, low range, 250 µl
161-0311 Biotinylated Standards, high range, 250 µl
161-0319 Biotinylated Standards, broad range, 250 µl

Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547

Phone [phone]

LIT396 Rev F

Cytochrome c

Lentil lectin

Human hemoglobin C

Human hemoglobin A

Equine myoglobin

Equine myoglobin minor band

Human carbonic anhydrase

Bovine carbonic anhydrase

β-Lactoglobulin B

Phycocyanin

Fig. 1. IEF Standards for accurate pI calibration of native IEF gels. A. 5 µl of the standards were run on a horizontal

polyacrylamide IEF gel and stained with Coomassie blue R-250 dye and crocein scarlet. B. 5 µl of the standards were
run on a horizontal polyacrylamide IEF gel and left unstained.

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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 deion­ized, destilled water as a diluent.

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56

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 repro­ducible 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 ana­lytical acrylamide or agarose isoelectric focus­ing 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.