Bio-Rad DEAE 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 see Table 2
Regeneration buffer 2 M guanidine HCl in application
buffer, or 1.5 M NaSCN
Buchner funnel
Chromatography column
General Instructions
1. Prepare the appropriate buffer using the information
given in Table 1. Accurate buffer preparation is
essential for optimum IgG recovery. See Optimizing
the Separation Conditions.
Table 1. Buffers for Purification of IgG from Serum on DEAE Affi-
Gel Blue gel
Species Application Buffer
Rabbit 20 mM Tris-HCl, pH 8.0, 25 mM NaCl,
0.02% NaN
3
Sheep as for rabbit
Goat as for rabbit
Human* 20 mM K
2
HPO
4
, pH 8.0, 0.02% NaN
3
Mouse 20 mM Tris-HCl, pH 7.2, 25-50 mM NaCl
These buffers are recommended starting points. Minor changes may be
needed. See Optimizing the Separation Conditions.
*KH
2
PO
4
is used to prepare this buffer. The pH is adjusted with KOH.
3
superior method of obtaining serum fractions
uncontaminated by albumin.
Product Description
Matrix Bio Gel A-5m agarose gel
Particle size 150-300 µm (50-100 mesh)
Shipping medium 0.01M Tris, pH 8, 0.15 M NaCl,
0.04% NaN
3
Functional Groups Cibacron blue and diethylaminoethyl
Typical flow rate* 15-25 cm/hr
Pressure limit 15 psi
Capacity**
Serum 0.2-1 ml of serum/ml of gel
Recovery of IgG >55%
Recovery of Albumin >90%
Removal of protease 100%
Stability
pH 2-11
Organic solvents alcohols
Temperature not autoclavable
Storage 1 year at 4 °C, in 0.02% NaN
3
or
other preservative
*Flow rate determined using a 1.5 x 20 cm column, and a hydrostatic
pressure of 1:1.
**The capacity for rabbit serum and human serum is lot to lot dependent.
It is determined on every lot, and provided on the label.
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