Antibody Problems

patingsadagat via methods%40net.bio.net (by arnigambe from gmail.com)
Tue Jan 8 20:41:08 EST 2008



> Right. I was about to ask if your protein had any homology to tubulin, and
> could be inducing a cross-reacting antibody response

The protein I'm studying does not have any homology to tubulin.
>
> I think the fact that you have 6his-tagged antigen makes affinity
> purification a lot easier, if you're prepared to expend some resin on it:

I performed an experiment exploiting the fact that my recombinant
protein is 6his-tagged using Reacti-Gel from Pierce. However, I was
not able to get enough antibodies. I guess I can try this experiment
one more time using tubulin-blocked serum solutions.

>Any chance you can complain to the antibody company and get
>them to do it again with a rabbit that isn't already making tubulin
>antibodies

I'm not sure if I can get them to do it all over again. It takes too
much time and I am trying to beat a deadline.

Thank you so much for your helpful comments and suggestions.


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