Helen-
I agree with Derek's assessment that blue sepharose is good to 'deplete' albumin, but not eliminate it. It often takes more than one pass over a column. Especially since you refer to conditioned media as the starting material. Media is often supplemented with 5 or even 10% fetal calf serum, which contains a "ton" of albumin. To deal with this, we actually switched the cells to serum-free media for their last several hours before examining media for our protein of interest, and still found it necessary to use more than one pass over blue sepharose. Blue sepharose does work, but it can't work miracles in the face of mega-quantities of albumin.
Tom
____________________________
Thomas T. Andersen, Ph.D.
Assistant Dean
Albany Medical College
Albany, NY 12208
518 262-5253
518 262-5183 fax
anderst@mail.amc.edu
>>> "Helen Kim, Ph.D." <Helen.Kim@ccc.uab.edu> 02/16/01 05:42PM >>>
Hi All,
Has anyone had experience removing albumin with Blue-sepharose? I
know in principle it can be done, and I've read lots of papers where people
blithely say they did with the blue sepharose. I recommended the
procedure to a grad student dealing with blob of albumin in his 2-D gels
of conditioned media. He bought some, tried it (I don't know exactly what
he did), and he says "it didn't work." He's very bright, though, so I trust
he went through reasonable efforts, but I thought I would check with you
all, since there's no point re-inventing the wheel.
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