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?
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