SPDP-C6-NHS ester

SPDP 试剂的胺反应部分是 N-羟基琥珀酰亚胺（NHS）酯。反应最通常在 pH 7-8 的磷酸盐、碳酸盐/碳酸氢盐或硼酸盐缓冲液中进行。可以使用其他不包含伯胺（或硫醇或二硫化物还原剂–参见巯基反应性的讨论）的缓冲剂。pH 值越高，反应和水解降解的速率越大；例如，NHS 酯的半衰期在 pH 值为 7 时为数小时，而在 pH 值为 9 时不到 10 分钟。SPDP 和 LC-SPDP 等 NHS 酯试剂的水溶性有限，在将其添加到反应混合物之前，必须先将其溶于有机溶剂中。诸如 Sulfo-LC-SPDP 的 Sulfo-NHS-酯试剂是水溶性的，可以直接添加到水性反应混合物中。 特点和优势： 反应性基团：NHS 酯和吡啶基二硫醇 对以下分子具有反应性：氨基和巯基 与游离巯基反应时释放出可检测的副产物；通过测量 343 nm 处吡啶-2-硫酮的释放，可以轻松地跟踪反应进度 间隔臂中的二硫键在pH 8.5条件下易于被10-50mM DTT或TCEP裂解 减少 SDS-PAGE 上样缓冲液，也易于使间隔臂裂解 使用可裂解交联剂，能够实现交联产物的分离 水不溶性（先溶于 DMF 或 DMSO） SPDP 交联剂可透过膜，因此可以在细胞内部进行交联 与其他品种的 SPDP 型试剂进行比较，包括聚乙二醇化形式 The amine-reactive portion of SPDP reagents is the N-hydroxysuccinimide (NHS) ester. Reactions are most commonly performed in phosphate, carbonate/bicarbonate, or borate buffers at pH 7-8. Other buffers can be used provided they do not contain primary amines (or thiols or disulfide reducing reagents – see discussion of the sulfhydryl reactivity). The rate of reaction and degradation by hydrolysis increases with increasing pH; for example, the half-life of the NHS ester is several hours at pH 7 and less than 10 minutes at pH 9. NHS-ester reagents like SPDP and LC-SPDP have limited aqueous solubility and must be dissolved in organic solvent before adding them to a reaction mixture. Sulfo-NHS-ester reagents like Sulfo-LC-SPDP are water-soluble and can be added directly to aqueous reaction mixtures. Features and advantages: Reactive groups: NHS ester and pyridyldithiol Reactive towards: amino and sulfhydryl groups Releases a detectable by-product when reacted with a free sulfhydryl group; by measuring the release of pyridine-2-thione at 343 nm, the reaction can be easily followed Disulfide bond in the spacer arm is readily cleaved by 10-50 mM DTT or TCEP at pH 8.5 Spacer arm is also easily cleaved using reducing SDS-PAGE sample loading buffer Cleavable crosslinker allows separation of crosslinked products Water-insoluble (dissolve first in DMF or DMSO) SPDP crosslinker is membrane permeable, so crosslinking can be done inside the cells Compare to other varieties of SPDP-type reagents, including pegylated forms

No numbering system information yet

No physical and chemical property information yet

No safety information yet

No production method and use information yet

No upstream information yet

No downstream information yet

No related product information yet