Bio-Rad DEAE Affi-Gel Blue Gel - Manual (Page 6)

2. What is the purpose of the regeneration buffer wash the first time the
column is used?
For new preparations of gel, this initial wash will elute residual blue dye
which might otherwise be eluted in serum protein fractions.
3. What are typical flow rates?
0.8-1.5 ml/min.
4. What is the shelf life of the Econo-Pac serum IgG purification kit?
The shelf life of the Econo-Pac serum IgG purification kit is 1 year at 4 °C.
The unreconstituted buffers are good for at least 1 year when stored at room
temperature in a tightly sealed bottle.
Section 2
Econo-Pac Serum IgG Purification Columns
Catalog Number 732-2026
2.1 Introduction
Econo-Pac serum IgG purification columns are Econo-Pac 10 chromatog-
raphy columns packed with 10 ml of DEAE Affi-Gel blue gel with an upper frit
above the gel bed. DEAE Affi-Gel blue gel consists of Cibacron
® Blue F3GA
dye coupled to specially prepared DEAE Bio-Gel ® A gel. Chromatography on
DEAE Affi-Gel blue agarose gel produces a purified IgG fraction with only
residual contamination from transferrin. After the IgG has been eluted, additional
serum fractions can be obtained from the column by elution with an ionic
strength gradient.
The Econo-Pac serum IgG purification columns provide a rapid and con-
venient way to purify IgG from serum.
2.2 Buffer Consideration
Prewash
When using an Econo-Pac serum IgG purification column for the first time,
a prewash is required to elute from the column residual dye which might oth-
erwise be eluted in serum protein fractions. Wash the column with 40 ml (fill the
column twice) of either:
• 1.5 M sodium thicyanate, or
• 0.10 M acetic acid, pH 3.0, containing 1.4 M NaCl and 40% (v/v)
isopropanol.
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