Bio-Rad Affi-Gel Protein A Media User Manual
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4. Shelf life to the Econo-Pac protein A kit:
The shelf life of the Econo-Pac protein A column is 1 year at
4 °C. The unreconstituted buffers are good for at least 1 year
when stored at room temperature. Reconstituted buffers can
be stored at 4 °C for approximately 2 weeks.
5. Purification of IgG from serum or tissue culture fluids:
In addition to ascites, serum and tissue culture samples are
suitable as samples for the Econo-Pac protein A kit. Serum
samples will require the same sample preparation and purifi-
cation procedures as ascites samples (as outlined in this manu-
al).
Tissue culture fluids should be concentrated to approximately
5 mg immunoglobulin/ml and then processed on an Econo-
Pac 10DG desalting column as described in this manual (or
diluted one to one with binding buffer). The purification pro-
cedure for the concentrated tissue culture fluid will then be
identical to the ascites procedure.
6. Purification of IgG other than mouse:
Human and rabbit IgG from serum (Bio-Radiations No. 53)
have been purified successfully with Affi-Gel protein A gel.
The protocol in this manual can be used without modification.
7. Purification of IgM with the Econo-Pac protein A kit:
Protein A will bind approximately 50% of all IgMs. The
purification of IgM using the Econo-Pac protein A kit will
therefore vary with each particular IgM sample.
Section 2
Econo-Pac Protein A Columns
2.1 Introduction
Bio-Rad’s Econo-Pac protein A columns are Econo-Pac 10
chromatography columns packed with 2 ml of Affi-Gel protein A gel
with an upper frit above the gel bed. Affi-Gel protein A consists of
purified protein A coupled to crosslinked agarose beads via stable
amide bonds. When used with the patented MAPS II buffer sys-
tem, each column has a capacity of 10-14 mg mouse IgG
1
.
The Econo-Pac protein A columns provide a rapid and conve-
nient way to purify monoclonal antibodies from small amounts of
ascites fluid. Binding, elution, and regeneration steps can be com-
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