Greetings you all,
I am trying to conjugate "CMDSK(phospho-S S)(phospho-S)GWGM" phosphopetide (11-mer) to a His-tagged 43 kDa recombinat protein. Could you please share your protocol(s) or provide me advices in this task.
Your help is much appreciated !
Regards,
Baki
Hi,
I assume you want to use the C-terminal cysteine to conjugate the phosphopeptide to the protein?
If so, you need to make sure that all cysteines involved are reduced (i.e. free syulfhydryls), bring the two together and perform some kind of re-oxidation.
Do you know how many cysteines are in your recombinant protein of interest? And whether these are involved in disulfide bonds that are essential for protein conformation?
If the protein has an odd number of cysteines, try to mildly reduce by using TCEP prior to reoxidation in the presence of your phosphopeptide.
Hope this helps.
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