The presentation of SULFO-SMCC

Water Acrid SMCC; Sulfo-SMCC X-linker; SSMCC; S-SMCC; Sulfosuccinimidyl-4-N-maleimidomethyl-cyclohexane-1-carboxylate; Sulfo SMCC and its water-soluble analog Sulfo-SMCC are heterobifunctional crosslinkers that accommodate N-hydroxysuccinimide (NHS) ester and maleimide groups that acquiesce covalent alliance of amine- and sulfhydryl-containing molecules. NHS esters acknowledge with primary amines at pH 7-9 to anatomy amide bonds, while maleimides acknowledge with sulfhydryl groups at pH 6.5-7.5 to anatomy abiding thioether bonds. In aqueous solutions, NHS ester hydrolytic abasement is a aggressive acknowledgment whose amount increases with pH. The maleimide accumulation is added abiding than the NHS-ester accumulation but will boring hydrolyze and loses its
reaction specificity for sulfhydryls at pH ethics > 7.5. For these reasons, conjugations with these crosslinkers are usually
performed at pH 7.2-7.5, with the NHS-ester (amine-targeted) reacted afore or accompanying with the maleimide (sulfhydryltargeted) reaction. The cyclohexane ring in the spacer arm of these reagents decreases the amount of hydrolysis of the maleimide accumulation compared to agnate reagents that do not accommodate this ring.
1This affection enables proteins that accept been maleimide-activated with SMCC or Sulfo-SMCC to be lyophilized and stored for after alliance to a sulfhydryl-containing molecule. Abounding maleimideactivated protein articles are produced in this address (see Related Products).
SMCC and Sulfo-SMCC are about acclimated to adapt antibody-enzyme and hapten-carrier protein conjugates in a two-step acknowledgment scheme. First, the amine-containing protein is reacted with a several-fold molar balance of the crosslinker, followed by abatement of balance (nonreacted) reagent by desalting or dialysis; finally, the sulfhydryl-containing atom is added to acknowledge with the maleimide groups already absorbed to the aboriginal protein. Sulfo-SMCC is acrid in baptize and abounding added aqueous buffers to about 10 mM, although solubility decreases with accretion alkali concentration. SMCC is not anon water-soluble and have to be attenuated in an amoebic bread-and-butter such as dimethylsulfoxide (DMSO) or dimethylformamide (DMF); consecutive concoction into aqueous acknowledgment absorber is about possible, and a lot of protein reactants will abide acrid if the final absorption of amoebic bread-and-butter is beneath than 10%.
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