Amplex® Red Phosphatidylcholine-Specific Phospholipase C Assay Kit

Catalog number: A12218

Molecular Probes™  Related applications: Protein Assays and Analysis

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Description

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.

See our complete line of Fluorescence Microplate assays.

• Detects phospholipase-C activity levels as low as 0.2 mU/mL
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference

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.

Because resorufin has absorption and fluorescence emission maxima of approximately 571 nm and 585 nm, respectively, there is little interference from autofluorescence in most biological samples.

Use Amplex® Red Assays for a Broad Range of Investigations
A wide variety of validated Amplex® Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex® UltraRed Reagent (Cat. No. A36006), a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex® Red/UltraRed Stop Reagent (Cat. No. A33855). The Amplex® Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least three hours. Custom assay design and packaging are also available.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

For Use With (Equipment): Microplate Reader, Fluorometer
Detection Method: Fluorescent
Format: 96-well plate, Cuvettes
Label or Dye: Amplex® Red
Product Size: 500 assays
Shipping Condition: Room Temperature

Contents & storage

Store in freezer (-5 to -30°C) and protect from light.

Documents

Manuals & protocols