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eliminating albumin from samples for western blotting


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#1 Paola PC

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Posted 01 August 2002 - 08:48 AM

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
Thanks

#2 milanoj

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Posted 01 August 2002 - 10:11 AM

Paola,
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.




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