Broadpharm代理商-BP Fluor 594 C5

做了这么多的 WB，你知道化学发光和荧光可以一起检测吗？

，一组使用 NaHS 处理，一组则不作处理；然后用荧光马来酰亚胺染料 Alexa Fluor 488 C5 maleimide（下文简称MAL，Invitrogen，A10254）标记半胱氨酸上的硫醇基团，使正常的半胱氨酸通过 -S- 与 MAL 连接，而硫巯基化的半胱氨酸通过二硫键 -S-S- 与 MAL 连接，如下图： 图 1. 文献 Fig. 6G，硫巯基化分析步骤 接下来加入 DTT，硫巯基化蛋白上特有的二硫键就会被解开，导致荧光 MAL 分子脱落，荧光信号减弱。所以，荧光信号发生变化就验证

Site-Specific Labeling of Proteins for Single-Molecule FRET Measurements Using Genetically Encoded Ketone Functionalities

to the amber stop codon TAG. Proteins were purified with high yield and subsequently labeled with the hydroxylamine derivative of Alexa Fluor� 488 functioning as a fluorescent donor dye. The proteins were then labeled via maleimide coupling chemistry

Overview of Biacore Systems and Their Applications

Myszka, D.G., He, X., Dembo, M., and Morton, T.A. 1998. Extending the range of rate constants available from Biacore: Interpreting mass transport‐influenced binding data. Biophys. J. 75:583‐594.