CNBR Digestion on SDS-PAGE gels Slices??

Tom Thatcher ttha at uhura.cc.rochester.edu
Thu Oct 5 13:10:25 EST 1995


In <Roberts_Daniel_g.PRILVMS3-0510950755460001 at riversend.bms.com> Roberts_Daniel_g.PRILVMS3 at msmail.bms.com (Dan Roberts) writes:

>Does anyone out there know of any methods for doing a CNBr
>digestion directly on an SDS-PAGE gel slice??.>thanks.>Dan


I realize this is going to sound like a shameless plug, but I did it
in one of my papers, and you could get the reference I used from it. 
Basically you soak the gel slice in an excess volume of 20 mg/ml
CNBr in 70% formic acid, overnight at room temp in the dark; then
collect the liquid and lyophilize it; dissolve the dried protein 
in water and lyophilize again (to get rid of trace CNBr and Formic Acid).

The paper is Thatcher,TH and Gorovsky, MA.  
Experimental Cell Research.  209(2):261-70, 1993 Dec.


-- 
Tom Thatcher                          | You can give a PC to a Homo habilis,
University of Rochester Cancer Center | and he'll use it, but he'll use it
ttha at uhura.cc.rochester.edu           | to crack nuts.



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