Secreted Protein Concentration

Guy Hermans ghermans at luc.ac.be
Fri Apr 24 02:27:47 EST 1998


In article <3.0.32.19980423085502.006a5d1c at itsa.ucsf.edu>,
fauss at ITSA.UCSF.EDU ("Donald J. Fauss") wrote:

> 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

How big is your protein? You may try to dialyse the conditioned supernate
to PBS, and spin the volume down one of those concentrators (with 10,000
Dalton cut-off filter)

Hope this helps,

Guy

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Guy Hermans, PhD student
MS research Unit             Tel 0032 (0) 11/26.92.07
Dr. L. Willems-Institute     Fax 0032 (0) 11/26.92.09
University Campus            E-mail ghermans at luc.ac.be
B-3590 Diepenbeek
Belgium
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