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problem of specific Thr phosphorylation detection - (Jan/04/2007 )

Hi:

I posted similar topic in protein and proteomic section. I have trouble to detect specifc Thr phosphorylation by in-house antibody.
This antibody was affinity purified by phosphopeptide. My treatment is working by blotting with other antibodies, but, i couldn't pick up any signals by this in-house antibody.Any suggestions?

-yxz98-

QUOTE (yxz98 @ Jan 5 2007, 08:33 AM)
Hi:

I posted similar topic in protein and proteomic section. I have trouble to detect specifc Thr phosphorylation by in-house antibody.
This antibody was affinity purified by phosphopeptide. My treatment is working by blotting with other antibodies, but, i couldn't pick up any signals by this in-house antibody.Any suggestions?


if it is selfmade, how did you qualify specificity of your Ab?
we once immunized six rabbits with same immunogene and succeeded with a well working Ab only in one case; one showed nothing, all others were mediocre to bad

-The Bearer-