I have deposited a layer of thiolate nanoparticles onto the substrate surface. Used dye and verified that SH group is active. Then I tried to use a maleimide/NHS ester crosslinker, but it seems like it is not working. I am dissolving SMCC in DMF and adding it to reagent alcohol to reach 4mM and soaked the substrate in for one hour for the maleimide to react with thiol. I don't think the reaction is successful.
Hello Johnny,
Try the following two things:
Use DMAc (Dimethylacetamide) instead of DMF. Alternatively, purge DMF with nitrogen and keep it under dehydration sieves.
Add DIPEA (N,N-Diisopropylethylamine) to the maleimide-thiol reaction, using a ratio of 3 moles equivalent to SH groups.
Checking out this publication Article Site-Specific, Stoichiometric-Controlled, PEGylated Conjugat...
Good luck,
Mateusz
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