Re: PepSyn: Phospho peptides for antibodies?
Angela c. Murphy (acmurphy@helix.nih.gov)
Tue, 16 Jun 1998 15:36:15 -0400 (EDT)
Phosphopeptides can be used to raise antibodies if they are conjugated to
a protein like KLH. I synthesized 3 peptides for one of our postdocs
(phosphorylation sites of acanthamoeba myosin I isoforms). I also
synthesized the unphosphorylated peptides. The postdoc conjugated one of
the peptides using glutaraldehyde, and the other 2 using an N-terminal
cysteine. This was done both with the phosphorylated and unphosphorylated
peptides. These peptides were all HPLC purified, to make sure that there
was no unphosphorylated peptide contaminating the phosphopeptide. The
postdoc got very nice, specific antibodies to the phosphopeptides, which
did not cross-react with the unphosphorylated peptides. He went on to use
the antibodies to study the localization of the phosphoproteins in intact
acanthamoeba.
Hope this helps.
Angela Murphy
On Tue, 16 Jun 1998, Lisa Bibbs wrote:
> We don't have any experience using phospho peptides for antibodies. Is it
> recommended?
>
> Thanks for the help as usual,
>
> Lisa
>
>
*******************************************
Angela C. Murphy, Chemist
Lab. of Cell Biology, NHLBI, NIH
3 Center Drive, MSC 0301, Rm. B1-22
9000 Rockville Pike
Bethesda, MD 20892-0301 USA
tel.: (301) 496-2324
fax: (301) 402-1519
email: acmurphy@helix.nih.gov
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