Bio-Rad CM Affi-Gel Blue Gel User Manual

CM Affi-Gel®Blue Gel

Instruction Manual

Catalog Number

153-7304

Introduction

o

NH

2

o

NH

SO2ONa

NH

N

N

N

O

Crosslinked

Agarose

NH

R

1

R

2

R1 = H or SO2ONa
R

2

= SO2ONa or H

CM Affi-Gel blue gel is a bifunctional affinity/ion exchange
chromatography matrix prepared by coupling Cibacron blue
F3GA and carboxymethyl (CM) groups to Bio-Gel®A-5m
crosslinked agarose gel. The Cibacron blue functions as an ionic,
hydrophobic, or sterically active binding site for proteins with
dinucleotide folds, such as albumin.

Cibacron blue coupled to agarose.

The carboxymethyl functional group functions as a cation
exchanger and will bind proteins with isoelectric points lower
than the pH of the mobile phase. (At pHs below 4.5 the car­boxymethyl group will be protonated and will not function as an
ion exchanger.) This bifunctionality allows CM Affi-Gel blue
gel to bind both albumin and protease from serum. Dialysis of

1

serum is not necessary prior to chromatography on this gel.
Albumin and serum protease are bound strongly enough that a
high concentration of salt or chaotropic reagent is required to
elute these components. Chromatography on CM Affi-Gel blue
gel provides a convenient initial step in the purification of serum
proteins.

Product Description

Matrix Bio Gel A-5m agarose gel
Particle size 150–300 µm (50-100 mesh)
Shipping medium 50 mM KCl with 0.04% NaN
Functional groups Cibacron blue and carboxymethyl
Flow rate 15–25 cm/hr
Pressure limit 15 psi
Capacity

Serum 2 to 6 ml of gel/ml of serum (lot

Stability

pH 2–11
Organic solvents alcohols
Temperature autoclavable

Storage 1 year at 4 °C, in 0.02% NaN

dependent)

other preservative

3

Material required but not supplied

Pre-wash buffer 0.1 M acetic acid, pH 3, 1.4 M NaCl,

Running buffer 10 mM K

Regeneration buffer 2 M guanidine HCl or 1.5 M NaSCN
Buchner funnel
Chromatography column

40% isopropanol

NaCl, 0.02% NaN

HPO4, pH 7.25, 0.15 M

2

,

3

General Instructions

1. Prepare CM Affi-Gel blue gel by washing it on a Buchner
funnel, or in a column, with 5 bed volumes of pre-wash
buffer, followed by 7 bed volumes of deionized water. This
is required the first time the gel is used, or if the gel has been
stored for more than 1 week since the previous use. Small
amounts of the blue dye may appear in the alcohol wash.
After prewash, equilibrate the column with 3 bed volumes
of running buffer.

The capacity of the gel is lot dependent, and will range
between 2–6 ml of gel per ml of serum. The capacity for

or

3

human and rabbit serum for a specific lot of gel is printed on
the label on the bottle.

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32

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