Bio-Rad Affi-Gel Protein A Media - Manual (Page 9)

5. Elute the IgG with 10 ml of elution buffer. Elute the column
with an additional 20 ml of buffer to insure total removal of
IgG. In most cases, the majority of the IgG will elute in the first
10 ml. Prolonged exposure to acid pH should be avoided.
Neutralize immediately after elution. The Econo-Pac 10DG
desalting columns can be used to rapidly neutralize, desalt and
buffer exchange the eluate into a suitable buffer (e.g., PBS).
[Note: the use of the Econo-Pac 10 DG desalting columns effec-
tively eliminates the need for lengthy dialysis procedures required
when samples are to be analyzed by SDS-PAGE.]
6. Wash the column with 10 ml of regeneration buffer.
7. Wash the column with 10 ml of binding buffer for the next chro-
matography cycle or store in PBS containing 0.05% sodium
azide at 4 °C.
2.5 Column Performance
Packed gel Affi-Gel protein A
Bed volume 2.0 ml
Reservoir volume 30 ml
Colume capacity 10-14 mg IgG
1
Recommended sample
volume
Maximum 2.0 ml ascites or serum
Packing buffer 10 mM sodium phosphate, 150
mM NaC1, pH 7.0, with 0.02%
sodium azide
Flow rate range 0.5-1.5 ml/min
Column material Polypropylene
Frit material Nominal 35 µm porous, polyethy-
lene
pH range pH 2-11
Recommended storage 4 °C, with PBS containing 0.02%
sodium azide
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