RE: Peptide competition Assay

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From:"Hewlett Bryan (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA>
To:"'histonet@Pathology.swmed.edu'" <histonet@Pathology.swmed.edu>, "'Carol Burden'" <caroltb@netscape.net>
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Date:Mon, 10 May 1999 08:38:52 -0400
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Carol,

This is a very easy procedure and one that we perform with all "in
house" polyclonal antibodies. Briefly, once we have established the
working dilution, we add a tenfold excess of the purified immunizing
peptide to a sample of the diluted primary. Leave overnight at 4C, spin
down hard the next day and use the supernatent as primary. There should
be NO staining of the item of interest.

Bryan

>----------
>From: 	Carol Burden[SMTP:caroltb@netscape.net]
>Sent: 	May 7, 1999 3:59 PM
>To: 	histonet@Pathology.swmed.edu
>Subject: 	Peptide competition Assay
>
>Hi ,
>I'm trying to detect an antibody that has been prepared by an investigator
>and
>is therefore largely uncharacterized.  We wish to verify the antibody by
>performing a peptide competition.  Has anyone done this before with tissue??
>
>Sincerely Thanks,
>Carol Caroltb@netscape.net
>
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