eliminating albumin from samples for western blotting - (Aug/01/2002 )
I am starting to study a protein which is secreted from cultured mammalian cell lines. I need to concentrate the conditioned medium to do some Westerns. The problem is that this way I have so much albumin from serum in my samples which makes a "blob" in the gel... How could I get rid of it? Is there any affinity chromatography I can use to absorb albumin?
my protein is about 30 KDa and pI is 6.66; albumin is 68KDa and pI is 5.8
Concentrate over a Centicon MWC 40,000 or 50,000 save the eluate, run on gel and see if your rid of the albumin contaminant.