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.
University of California - Davis