SMCC结构反应解释.docxVIP

SMCC 是一类含有 N-羟基琥珀酰亚胺(NHS)活性酯和马来酰亚胺的双功能偶联剂.可以将分别含有巯基和氨基的化合物键接在一起。 NHS 活性酯与伯胺在PH7-9 的环境形成酰胺键。马来酰胺与巯基在的环境下形成稳定的硫醚键。在水溶液中，NHS 活性酯的水解是（与氨基的反应）个竞争反应。马来酰胺比NHS 稳定，但是在PH 大于时，马来酰胺会慢慢水解，失去与巯基反应的特异性。因而， 在使用 SMCC 时通常是在的环境下进行，并且先让NHS 发生反应。 SMCC 结构里的环己烷环可以降低马来酰胺的水解速率。这使得蛋白质在用 SMCC 修饰之后可以冻干存放一段时间。很多蛋白质都选用该试剂来进行马来酰亚胺修饰。 用 SMCC 来制备抗体-酶或者半抗原作载体的蛋白质，经常采用两步合成法。首先，含有氨基的蛋白质与几倍的偶联剂反应，反应结束后通过脱盐柱或者透析的方法除掉没有反应玩的 SMCC。然后，再与含有巯基的蛋白质反应。在实际操作中要注意的是，SMCC 怕潮湿，存放时要和干燥剂一起存放。并且使用中从冰箱拿出来时要先在室外放置一段时间平衡温度，以免立刻开启，空气中水分遇冷凝结，破坏 SMCC 结构。 INSTRUCTIONS SMCC (succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxy late), 50 mg Molecular Weight: Spacer Arm: ? Net Mass Added: Storage: Upon receipt store desiccated at 4° C. Product is shipped at ambient temperature. Sulfo-SMCC (sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexan e-1-carboxylate), 1 g Sulfo-SMCC, 50 mg Sulfo-SMCC, No-Weigh Format, 8 × 2 mg microtubes Molecular Weight: Spacer Arm: ? Net Mass Added: CAS #: 92921-24-9 Storage: Upon receipt store desiccated at -20° C. Produc t is shipped at ambient temperature. Introduction SMCC and its water-soluble analog Sulfo-SMCC are heterobif unctional crosslinkers that contain N-hydroxysuccinimide (NHS) ester and maleimide groups that allow covalent conjugation of amine- and sulfhydryl-containing molecules. NHS esters react with primary amines at pH 7-9 to form amide bonds, while maleimides react with sulfhydryl groups at pH to form st able thioether bonds. In aqueous solutions, NHS ester hydroly tic degradation is a competing reaction whose rate increases with pH. The maleimide group is more stable than the NHS- ester group but will slowly hydrolyze and loses its reaction specificity for sulfhydryls at pH values . For these re asons, conjugations with these crosslinkers are usually perfor med at pH with the NHS-ester (amine-targeted) reacted befor e or simultaneous with the maleimide (sulfhydryl-targeted) rea ction. The cyclohexane ring in the