Protein inhibitor(s) during immunoprecipitation ?

ijiwaru at wheel.dcn.davis.ca.us ijiwaru at wheel.dcn.davis.ca.us
Fri Mar 15 13:29:14 EST 1996


In article <4i9jpq$l8 at mercury.hgmp.mrc.ac.uk>, pdezoysa at hgmp.mrc.ac.uk wrote:

> Dear Netters,
> 
> I would like to know if:
> A) any of you out there have encountered degradation of target protein
during immunoprecipitation (preliminary data indicates that the source of 
> the proteolytic activity is likely to be the rabbit serum itself) and; 
> B)also know which protease inhibitor (or coktail of inhibitors) could
inhibit this protease activity?
> 
> Thanks in advance,
> 
> Priyal de Zoysa.

Many, many moons ago when I used to do this sort of stuff, I remember
using a cocktail of N-ethylmaleimide, phenylmethylsulfonylfluoride,
aprotinin  with an option to add leupeptin.  This did a pretty good job of
keeping proteases in check, although if you are going to look for enzyme
activity, you may want to keep an eye on the N-EM.

Lyle Najita
Plant Pathology
University of California - Davis



More information about the Proteins mailing list