Posted 10 December 2004 - 09:41 AM
I have questions about generation of phosphospecific antibody. I am studying a protein, which I have already shown, can be phosphorilated. And I want to prepare polyclonal Ab specific for phosphorilated form of the protein. I can express my protein in E. coli and phosphorilate it in vitro. What is better way to get a phosphospecific antibody: develop antibody against phosphopeptide or develop antibody against phosphorilated forms of the protein, and than deplete against non-phosphorilated forms? I know that company normally recommend use a phosphopeptides, but can be second way better?
Many thanks in advance!
Posted 21 December 2004 - 06:12 AM
Alternatively, you could test phosphotyrosine, -serine, and -threonine antibodies which are available. one of them might work with your protein.
Posted 24 December 2004 - 04:00 PM
I also suggest that you ask a company to do it. I had a good experience about phospho-antibody. Polyclonal AB created by a phospho-peptide and double purified by using both phospho-peptide and non-phospho-peptide will yield a good result.
Posted 06 January 2005 - 02:29 PM