Bio-Rad CM Affi-Gel Blue Gel - Manual (Page 3)

Material required but not supplied
Pre-wash buffer 0.1 M acetic acid, pH 3, 1.4 M NaCl,
40% isopropanol
Running buffer 10 mM K
2
HPO
4
, pH 7.25, 0.15 M
NaCl, 0.02% NaN 3 ,
Regeneration buffer 2 M guanidine HCl or 1.5 M NaSCN
Buchner funnel
Chromatography column
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
human and rabbit serum for a specific lot of gel is printed on
the label on the bottle.
**
3 2
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
3
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
dependent)
Stability
pH 2–11
Organic solvents alcohols
Temperature autoclavable
Storage 1 year at 4 °C, in 0.02% NaN
3
or
other preservative
LIT308C 9/3/98 7:13 AM Page 2