In Allen et al (1993) BBA 1202, 135-142 we used Pharmacia's Blue-
Sepharose in 10 mM phosphate buffer, pH 6.8 containing 100 mM NaCl. Column was 1 x 14 cm run at room temp at 0.5 mL/min.
This will help to deplete the eluate of albumin, but by no means remove all of it. In fact, what we applied to this column was not serum but material which had already been subjected to either an electrophoretic or a chromatofocusing purification step. To start with serum, scale the column up considerably and plan to use it only a few times. There is so much albumin in serum that it is very difficult to remove all of it. Also, this assumes that what you want is the eluate and not the albumin: we never tried to recover the albumin from the column, only washed it away as best we could with high salt.
Tom Andersen
____________________________
Thomas T. Andersen, Ph.D.
Assistant Dean
Albany Medical College
Albany, NY 12208
518 262-5253
518 262-5183 fax
anderst@mail.amc.edu
>>> Low Teck Yew <medp9148@nus.edu.sg> 05/22/00 01:43AM >>>
Hi,
I would like to remove serum albumin from human serum with blue dextran or
cibracon blue. Can anybody show me the protocol for doing so?
Thanks in advance!
T.Y. Low
Biochemistry Department,
National University of Singapore.
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