how to conjugate the peptide to KLH

Bin Tang btang at POST.ITS.MCW.EDU
Tue Jun 30 14:52:11 EST 1998


Dear Dr Liu:

As far as I know, there are several different methods to coup the
synthetic peptide to carrier protein. Here is the protocol we used.

1.To a 15ml glass tube, add 5 mg KHL and 1ml borate buffer(PH10).Dissolve
  by gentle mixing on a stir plate, then add 10umol(5mg) synthetic 
  peptide.
2.Slowly add 0.5ml of freshly prepared 0.3% glutaraldehyde solution while
  stirring at room temperature for 2 hours.
3.Add 0.125ml of 1M glycine to block unreacted glutaraldehyde for 30 min.
4.Dialyze against orate buffer(PH8.5).

That's it. If you have any question, we can discuss it later.

Yours,

Dr. Tang,Bin
6/30/98

On 26 Jun 1998, Shang Xi Liu wrote:

> Date: 26 Jun 1998 09:10:47 -0700
> From: Shang Xi Liu <liusx at fimmu.edu.cn>
> To: immuno at net.bio.net
> Subject: how to conjugate the peptide to KLH
> 
> Hi, I'd like to know how to conjugate the synthetic hexapeptide KFSSFR to KLH as an immunogen. How can I dissolve the peptide and what crossing agent can be used and how can I determine the efficiency
> of conjugate?
> 
> 




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