Invitrogen™ Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit

Invitrogen™ Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit

The Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit provides a sensitive method for detecting phosphatidylcholine-specific phospholipase C (PC-PLC) activity using a fluorescence microplate reader or fluorometer.

In this enzyme-coupled assay, PC-PLC activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex™ Red reagent), a sensitive fluorogenic probe for hydrogen peroxide. PC-PLC converts phosphatidylcholine (lecithin) to phosphocholine and diacylglycerol. Alkaline phosphatase is added, which hydrolyzes phosphorylcholine to form choline. Choline is then oxidized by choline oxidase to form betaine and hydrogen peroxide. In the presence of horseradish peroxidase, the hydrogen peroxide reacts with Amplex™ Red reagent in a 1:1 stoichiometric ratio to generate the highly fluorescent product resorufin.