Non-isotopic Western Blots-Best

Valery Thompson valeryt at ag.arizona.edu
Fri Nov 18 11:54:00 EST 1994


In article <Cz9JCJ.4p9 at ncifcrf.gov>, reming at ncifcrf.gov (Mary Remington) says:
>
>  Our laboratory will be doing Western blots in a BSL 3 and it would
>  simplify our lives greatly to use a non-isotopic method.  I would
>  appreciate any thoughts on the kits on the market for this purpose.
>  Many thanks, Mary
>

In my lab we have been using chemiluminescent detection for western blots for
 about the past 1 1/2 years with good success.  We have used the kits from 
DuPont NEN and from Boehringer Mannheim.  Both work well and are about the
same price/blot.  We have not tried the Amersham kit only because it costs
more.  It takes a little more work to develop the procedure than using the
colorimetric reagents because of the sensitivity, but once the correct Ab
dilutions are found, the procedure is quite easy.  Also there is no problem
with waste disposal as when I-125 is used.  The sensitivity I have 
experienced is in the sub-nanogram range.  

I hope this helps.  Good luck.

Valery Thompson
Sr. Research Specialist
Muscle Biology Group 
University of Arizona



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