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PH6.5-7.5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7.5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PH7.2-7.5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-carboxylate),?50?mg??Molecular?Weight:?334.32?Spacer?Arm:?8.3???Net?Mass?Added:?219.09?Storage:?Upon?receipt?store?desiccated?at?4°?C.?Product?is?shipped?at?ambient?temperature.???Sulfo-SMCC?(sulfosuccinimidyl?4-[N-maleimidomethyl]cyclohexane-1-carboxylate),?1?g?Sulfo-SMCC,?50?mg?Sulfo-SMCC,?No-Weigh??Format,?8?×?2?mg?microtubes??Molecular?Weight:?436.37?Spacer?Arm:?8.3???Net?Mass?Added:?219.09?CAS?#:?92921-24-9?Storage:?Upon?receipt?store?desiccated?at?-20°?C.?Product?is?shipped?at?ambient?temperature.??Introduction?SMCC?and?its?water-soluble?analog?Sulfo-SMCC?are?heterobifunctional?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?6.5-7.5?to?form?stable?thioether?bonds.?In?aqueous?solutions,?NHS?ester?hydrolytic?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??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?before?or?simultaneous?with?the?maleimide?(sulfhydryl-targeted)?reaction.?The?cyclohexane?ring?in?th