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Supernatant for western - (Jul/11/2008 )


I'm attempting to collect supernatant from fibroblast cell culture for western blotting, so far unsuccessfully.
Does anybody have a protocol I can try?
I'm using 24-well cell culture plates and I would like to verify details as number of cell/well, minimal amount of medium, and collecting conditions.



What protein(s) are you looking at?

-Minnie Mouse-

Dilution will be your enemy in this, you need some way of concentrating the proteins present in the medium before you will be able to see them on a western.


My control is albumin
I'm looking for IL-1 and TGF