Sheep IgG's with Email
cchughes at uci.edu
Thu Jun 27 21:22:08 EST 1996
In article <-2406961545060001 at mcconvillec.can.bham.ac.uk>, @bham.ac.uk
(room 103) wrote:
> Hi folks,
> I'm having big problems using antibodies raised in sheep....
> when I develop Western blots of whole human cell extracts
> using the ECL system the background is so high even at dilutions of 5000
> that any specific signal is lost.However this is not a problem when
> testing the antibodies on Westerns against the fusion protein....
Sounds like you need to pre-absorb your ab against human cell lysates
(non-transfected) to get rid of cross-reacting abs. In theory the only
abs you should get back capable of binding to anything would be those
specific to the His tag. I should say that I've never actually tried
this! Maybe worth a try though -- better than the drain.
Dr. Chris C.W. Hughes
Dept. Mol. Biol. & Biochem.
Biol. Sci. II
Univ. California, Irvine cchughes at uci.edu
Irvine CA 92717 ph (714) 824 8771
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