Secreted Protein Concentration

Donald J. Fauss fauss at ITSA.UCSF.EDU
Thu Apr 23 10:57:45 EST 1998


I need help on concentrating secreted proteins from cultured primary cells.
I've tried 70% EtOH precipitation but that didn't seem to work. I'll try
AmSO4 precipitation next but I'm wondering if I should include say 0.1%
TX-100 to help with the more hydrophobic proteins (maybe just to keep them
from sticking to the wall of the centrifuge tube?). I'm collecting them in
plain DME so I know the high salt concentration might prove a problem.
Probably I could detect the proteins with silver staining but I'm thinking
I'd need to be able to detect them with Coomassie staining before I'd have
enough protein to hopefully isolate species of interest. I'd like to hear
from anybody who has been successful at this.

Don Fauss
email: fauss at itsa.ucsf.edu



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