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Sulfo-SMCC cross linking - how to increase reactivity of maleimide


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#1 Titoni

Titoni

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Posted 15 July 2010 - 07:06 AM

Hi
I have quite an old vial of SMCC reagent and decided to try it for cross linking of a small protein with an oligonucleotide containing thiol group. After overnight treatment of the protein I can see a good peak in MALDI-TOF (+220 of starting peak). After desalting the mixture on speen column and adding the oligo (pH 7.0 adjusted) nothing seems to be happening for already a couple of hours. I am wondering if the maleimide of the SMCC is still reactive or it's all gone. I would thought that succinimide will be desactivated first. Any advices how to push the reaction? (I am now speed vacuuming it to concentrate them a bit)
Thanks for any help




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