Temp. staining of SDS PAGE gels prior to renaturation??

Joe Mack mack at fcs260c.ncifcrf.gov
Sat Nov 13 23:01:10 EST 1993


In article <CGFpD7.ELs at usenet.ucs.indiana.edu> Bill Nowatzke <wnowatzk at ucs.indiana.edu> writes:
>I have been trying to follow a R. Burgess paper in which enzymatic
>activity was regained in a protein purification after crushing the
>suspected protein band from the gel. The problem that I have been having
>is the transient staing of the gel to identify the approx Mr prior to
>cutting out the band. The original paper recommends ppt the SDS in the
>gel with 0.2M KCl for 5 min and then destaining with water for up to one
>hr. When I tried this the portion of the gel below the dye front became
>very white, implying ppt, but the portion of the gel above the dye front
>only became slight opaque and I could only see very faint bands of heavly
>overloaded lanes. I was made aware of another transient staining tech
>using a 0.3M copper soln which I tried on the same gel with poor results
>(although this gel would not fully destaing from the KCl treatment and
>that probably interfered). 
>I would appreciate any advice/procedures that you may have successfully
>used or heard about. Thanks in advance!

I've heard of people soaking the gel in KOAc, where the protein becomes
opaque. Joe Mack mack at ncifcrf.gov



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