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This kits takes our stand alone assay for phospholipase A2, PLA2 (A10072) and combines the necessary Ez and lipids to run 2 complete 96 well microplate assays that monitor activity in purified enzyme preparations and cell lysates. The EnzChek Phospholipase A2 Kit provides enough reagents for 2 microplates, using 200 µl volumes in 96 well format to perform continuous fluorometric monitoring of PLA2 . This product offers an alternative to our (B7701), (bis-BODIPY® FL C11-PC) reagent, by providing an PLA2 selective substrate and one that is ratiometric, thereby lowering variations produced by instrumentation and assay conditions. Phospholipase A2 or PLA2 represents a family of enzymes that hydrolyze the sn-2 ester linkage of phospholipids. The activities of these enzymes play important roles in cardiovascular, inflammatory and nervous system disorders, and in cancers. The EnzChek® Phospholipase A2 substrate provides sensitive and continuous rapid real-time monitoring of PLA2 enzyme activities. This unique substrate is selective for PLA2 and can be used either in a intensity or ratiometric based detection mode. In intensity based detection mode the PLA2 activity is monitored by the intensity increase of a single wavelength at approximately 515 nm. In ratiometric analysis, which is based on the distinct fluorescence resonance energy transfer (FRET) emission of this substrate prior to and after cleavage, PLA2 is detected by changes in the emission intensity ratio at 515/575 nm with excitation at ≈ 460 nm. Either detection mode provides a simple method with low background, high sensitivity and high specificity for PLA2.
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