EnzChek® Direct Phospholipase C Assay Kit

Catalog number: E10215

Molecular Probes™  Related applications: Protein Assays and Analysis

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Description

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.
The EnzChek® Direct Phospholipase C Assay Kit provides a simple and robust method for monitoring PC-PLC activity. Each kit provides enough reagents for 10 microplates, using 200 µl volumes in 96 well format. 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 509 nm and 516 nm, 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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Color: Green
Readout: Kinetic
Technique: Fluorescence Intensity
Assay Type: Phospholipase Assay
Filter Type: FITC (Fluorescein) Filter
Label or Dye: BODIPY® FL
Product Size: 1 kit
Target Enzyme: Phosphatidylcholine-Specific Phospholipase C
Substrate Type: Phospholipase Substrate
Detection Method: Fluorescent
Substrate Properties: Chemical Substrate, Lipid-Based Substrate
For Use With (Equipment): Microplate Reader
Excitation⁄Emission (nm): 509⁄516
Shipping Condition: Room Temperature

Contents & storage

I kit Store kit at -<20°, protected from light, desiccated

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Manuals & protocols

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