To solubilize transferred proteins..HOW??

Bernard P. Murray, PhD bpmurray*STUFFER* at socrates.ucsf.edu
Wed Mar 10 22:01:40 EST 1999


In article <36E6A7BC.65B5F9E2 at mail.utexas.edu>, lnd at mail.utexas.edu wrote:

> Hi  Netters
> I have a question here......
> Can anyone tell me...if it's possible .... PRACTICALLY....... to get
> proteins into solution after being transferred on membrane (NC,
> PVDF....whatever..)?  Say....to increase protein purity and
> concentration.
> I never did that...but ...who knows..maybe you or your friends did?
> Say...to chop membrane with certain  protein band,  add
> SDS/tween/urea/TX....detergents in buffer....boil....incubate etc.etc.?
> Any suggestions would be highly appreciated.
> Regards
> Levan  Darjania
> Austin, TX

Someone can correct me if I'm wrong but I believe that
nitrocellulose is soluble in DMSO.  Depending on in just
what condition you want the protein afterwards this, or
another solvent, may be suitable.  I once looked into this
for electrophoretic purificatiion for immunisation but
later found the literature on immunising with actual
pieces of membrane (or PAGE gel).
     The major problem is that you don't get very much
protein by this technique.
     Good luck,
          Bernard
-- 
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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