blotting fixed gels

Peter pxpst2 at unixs.cis.pitt.edu
Sat Oct 17 20:53:57 EST 1998


In article <199810162238.QAA281468 at nestor.NMSU.Edu>, hroychow at NMSU.EDU
(Hiranya Roychowdhury) wrote:

>   Here is something that I know is possible: One can electrophorese
> out polypeptides fixed and stained (CBB-R) in acryl. gels. This is a routine
> method of obtaining antigens for some. 
>         So, it may be possible to equilibrate such gels in Towbin's and
> transfer the content to PVDF using the usual western transfer set up.

Possible...yes indeed.  BUT the yield will stink especially as the size of
the "peptide" increases.  When I try to retreive bands from fixed gels, I
simple apply 40 ma for 48 hrs and electroelute the bands into a dialysis
bag.  The big problem that I have found is removing the SDS since my goal
is to sequence it by MS MS

Peter

-- 
"don't you eat that yellow snow
       Watch out where the huskies go"  
                             FZ



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