Western Blotting Blues

Mad Scientist isl48143 at image.dk
Tue Dec 11 07:27:49 EST 2001


Do you use protease inhibitors in your storage buffer?? They usually do the
trick when it comes to degradation. Without inhibitors only short time with
the sample not being on ice could degrade your sample. I use "Complete"
inhibitor tablets from Roche. Besides that keep your samples on ice at ALL
times.

cheers
silas



"kalyan" <kalsriv at mailcity.com> wrote in message
news:e7b6a13.0112100859.499dda77 at posting.google.com...
> Hi All
>  I have been doing the western Blots with the proteins from mammalin
> cells. Interestingly, what I saw that after first western Blot, that I
> did immidiately I got good bands. But after the storage of protein
> extract for 7 to 8 days, I saw no bands at all, working with same
> antibody and conditions. Predictably it is because of the degradation
> of protein at the storage time. I kept it at -70 degree after the
> solublisation with 5X sample buffer after the lysis and extraction of
> the protein.
>
> Can someone suggest the best storage condition for the protein before
> and after the solublisation with sample buffer.





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