Thermo Fisher, E10215, EnzChek™ Direct Phospholipase C Assay Kit

Quantity: 1 Kit
Unit Size: Each
This kit provides continuous monitoring of phosphotidyl choline specific phospholipase C (PLC) activity in microplate based biochemical assays. The assay uses a PCL selective fluorogenic substrate that emits a bright green emission upon cleavage. PC-PLC plays a crucial role in many cell signaling pathways involved in apoptosis and cell survival, as well as diseases ranging from cancer to HIV1-7. The assay uses a glycerophospho-ethanolamine with a dye-labeled sn-2 acyl chain as a substrate for PC-PLC. Substrate cleavage by PC-PLC before the phosphate releases the dye-labeled diacylglycerol, which produces a positive fluorescence signal that may be measured continuously. The reaction product has absorption and fluorescence emission maxima of 509nm and 516nm, respectively. Using purified enzyme from Bacillus cereus, the assay can detect as little as 10 mU/mL PC-PLC after one hour at room temperature. The kit has been proven useful for characterizing PC-PLC inhibition, and since it offers a direct measurement, the potential for false positives in a compound screen is eliminated.
• Each kit provides enough reagents for 10 microplates, using 200μL volumes in 96 well formatEnzyme and Protein Activity Assays, Lipase and Phopholipase Activity, Proteins, Expression, Isolation and Analysis