Bio-Rad DEAE Affi-Gel Blue Gel - Manual (Page 8)

5. Elute the IgG with 20 ml of application buffer. For a more precise collection
method, collect fractions of volume approximately equivalent to that of the sam-
ple applied. Determine the absorbance at 280 nm of each fraction using a spec-
trophotometer and combine effluent tubes containing the unbound protein peak.
[For the isolation of other serum proteins, the column may be eluted with a
gradient of increasing NaCl concentration. A final saline concentration of
0.5 M will elute most serum proteins other than albumin.]
6. An optional step: Most of the bound albumin can be eluted by washing the
column with application buffer containing 1.4 M NaCl.
7. Regardless of whether or not step 6 is carried out, regenerate the column with
20 ml of regeneration buffer.
8. Wash the column with 30 ml of application buffer and continue with step 4
for the next chromatography cycle, or store the column at 4 °C in applica-
tion buffer containing 0.02% sodium azide.
2.5 Column Performance
Packed gel DEAE Affi-Gel blue gel
Bed volume 10 ml of packed gel
Total column volume 30 ml
Column serum capacity See label on column package for exact column
capacity
Packing buffer 10 mM sodium phosphate, 150 mM NaCl,
pH 7.0, with 0.02% sodium azide
Flow rate range 0.8-1.5 ml/min
Column material Polypropylene
Frit material Nominal 35 µm porous, polyethylene
Recommended storage 4 °C, in application buffer containing 0.02%
sodium azide.
Section 3
Product Information
Catalog
Number
Product Description
732-2027 Econo-Pac Serum IgG Purification Kit, includes Econo-Pac serum
IgG purification columns (5), Econo-Pac 10DG desalting columns
(5), human and rabbit application buffers, regeneration buffer.
732-2026 Econo-Pac Serum IgG Purification Columns, 5
153-7307 DEAE Affi-Gel Blue Gel, 100 ml
732-2010 Econo-Pac 10DG Desalting Columns, 30 columns
732-1015 Econo-Pac 10 Rack, 12 place
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