Western Blotting Blues

Mike Hanlon someone at asd.com
Fri Dec 14 06:58:50 EST 2001


Will all depend on the particular protein. Proteins that like a high salt
buffer for example tend to be less soluble at low temps. Yoy may have to
experiment if you cannot find someone with similar experience. You could
start by storing aliquots at different temps - say -20oC and 4oC.

Mike


"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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