390 通用MMP 活性试剂盒 荧光法、390 Generic MMP Activity Kit *Fluorimetric*详细如下: 390 Generic MMP Assay Kit 经过优化,可检测多种 MMP 的活性,包括 MMP-1、2、7、8、9、13、14、15、16 和 24。该试剂盒可用于检测生物样品中的通用 MMP 活性或使用纯化的 MMP 高通量筛选 MMP 诱导剂和抑制剂。该试剂盒提供 Mca/Dnp 荧光共振能量转移 (FRET) 肽作为 MMP 底物。在完整的 FRET 肽中,Mca 的荧光被 Dnp 淬灭。在被 MMP 切割成两个单独的片段后,Mca 的荧光被恢复,并且可以在激发/发射波长 = 330 nm/390 nm 下进行监测。检测以方便的 96 孔微孔板形式进行。 基质金属蛋白酶 (MMP) 属于能够消化细胞外基质成分的分泌型或膜相关蛋白家族。 Add test compounds and MMP diluent into microplate. The suggested volume of enzyme solution for a 96-well plate is 40 µL/well and test compound is 10 µL/well. Set up the following controls at the same time: Positive control contains MMP diluent without test compound. Inhibitor control contains MMP diluent and known MMPs inhibitor. Vehicle control contains MMP diluent and vehicle used to deliver test compound (e.g. DMSO). Test compound control contains assay buffer and test compound. Some test compounds have strong autofluorescence and may give false results. Substrate control contains assay buffer only. Using the assay buffer (Component E), bring the total volume of all controls to 50 µL. Optional: Pre-incubate the plate for 10 min. at assay temperature. Any temperature (the assay temperature) from room temperature to 37°C may be used, as long as the subsequent incubations are performed at the same temperature.