Protein acetylation - (Jul/05/2006 )

Hello!
I have a question concerning protein acetylation.
I want to analyze if a certain protein is acetylated or not. I'd like to do this by labeling cells with 3H- or 14C-sodium acetate. At the moment my problem is that I would like to have a negative control, a protein that is definitely not acetylated. In the literature I found up to now nothing. People use negative controls only in in vitro assays.
Maybe you can help me, and tell me a protein that is not acetylated!?

Thanks for your help!

thomas

-mercapto-

This isn't answering your question but do you know if the commercially available anti-acetyl antibodies work well. (I think Upstate/Cell signalling technologies make them).

Best wishes,
Ceri

-Ceri-

QUOTE (Ceri @ Jul 6 2006, 12:13 PM)

This isn't answering your question but do you know if the commercially available anti-acetyl antibodies work well. (I think Upstate/Cell signalling technologies make them).

Best wishes,
Ceri

Yes I have. But we get no signal with our protein, even if overexpressed and purified by IP. So we can make no real conclusion from this. Maybe were under the detection level, maybe the specific acetylated sequence in our protein is not recognized.

-mercapto-

Thinking about your negative control. If you can detect acetylation with your labelling method maybe you could then treat with a deacetylase to show the protein's definitely acetylated.

Ceri

-Ceri-