Hi,
I have carboxylated beads and I want to covalently attach a protein with the thiol group of a cysteine. Can I use SMCC chemistry for this? Are there linkers that will make this easier for me?
SMCC has a succinimidyl ester for reacting with amino groups and a maleimide for reacting with thiols. This would be a suitable crosslinker if your beads had primary amino group functionality.
If you first reduce the carboxylate groups to aldehydes, you could use BMPH to add a thiol-reactive crosslinker:
https://www.thermofisher.com/order/catalog/product/22297?SID=srch-srp-22297
Protein has NH2(from lysines or NH2 end , NH2 can connect to COOH bead!
The trick with crosslinkers in this situation is to avoid crosslinking the protein to itself, either intra- or intermolecularly. You could use a carbodiimide like EDC to crosslink amino groups in the protein to the carboxyl groups on the beads by adding the carbodiimide to the mixture of beads and protein, but you would likely end up crosslinking the protein to itself to some extent. It is better to introduce the protein-reactive functional group onto the beads first, wash out excess crosslinker, then add the protein to the functionalized beads, thus avoiding any protein-protein crosslinking.
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