Thermo Scientific Pierce LC-SMCC is an amine-to-sulfhydryl crosslinker that contains NHS-ester and maleimide reactive groups at opposite ends of a long cyclohexane-stabilized spacer arm （16.2 angstroms）.
LC-SMCC is a non-cleavable and membrane permeable crosslinker. It contains an amine-reactive N-hydroxysuccinimide （NHS ester） and a sulfhydryl-reactive maleimide group. NHS esters react with primary amines at pH 7-9 to form stable amide bonds. Maleimides react with sulfhydryl groups at pH 6.5-7.5 to form stable thioether bonds. The maleimide groups of LC-SMCC and are unusually stable up to pH 7.5 because of the cyclohexane bridge in the spacer arm.
LC-SMCC is often used for the preparation of antibody-enzyme and hapten-carrier conjugates. In this type of conjugation, the NHS ester is reacted first with the antibody, excess crosslinking reagent removed and then the sulfhydryl-containing enzyme molecule is added. This two-step reaction scheme results in formation of specific antibody-enzyme conjugates.
LC-SMCC is a reagent of choice for creating stable maleimide-activated carrier proteins that will spontaneously react with sulfhydryls. In this application, the NHS ester of LC-SMCC is reacted with lysine residues on the carrier protein, converting them to reactive maleimides. These relatively stable maleimide-activated intermediates may be lyophilized and stored for later conjugation to a hapten.
Highlights:
Amine reactive NHS ester crosslinks rapidly with primary amine-containing molecule
Sulfhydryl-reactive maleimide reacts with cysteine residues to yield specific conjugates
High purity, crystalline LC-SMCC can be used to create high-purity maleimide-activated derivatives
Cyclohexane bridge confers added stability to the maleimide group making LC-SMCC an ideal crosslinking agent for maleimide activation of proteins. Maleimide groups are stable for 64 hours in 0.1 M sodium phosphate buffer, pH 7 at 4°C。
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