peptide conjugation

Michael Jobling (m.jobling@pgrad.unimelb.edu.au)
Wed, 20 Jan 1999 11:15:19 +1100

Hi everybody.
We are trying to conjugate KLH to a peptide (via a free cysteine). The
problem is that our peptide has two cysteine residues which form a
disulphide bond quite readily. After reduction by DTT, the disulphide
reforms during dialysis to remove the DTT from the reaction mixture. Apart
from running a column or 'quicker' method of removing the DTT, so the
disulphide doesn't have a chance to reform, does anybody have a suggestion
as to how we might be able to maintain a free S-H to add the KLH to.
Thanks in advance

Michael Jobling
Dept. of Pathology
Uni. of Melbourne
m.jobling@pathology.unimelb.edu.au