Thermo Fisher Scientific 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.

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

Assay Principle

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.

1. PC-PLC converts phosphatidylcholine (lecithin) to phosphocholine and diacylglycerol.
2. Alkaline phosphatase hydrolyzes phosphorylcholine to form choline.
3. Choline is oxidized by choline oxidase to form betaine and hydrogen peroxide.
4. In the presence of horseradish peroxidase (HRP), hydrogen peroxide reacts with Amplex™ Red reagent to generate the highly fluorescent product resorufin.

Resorufin exhibits absorption and fluorescence emission maxima at approximately 571 nm and 585 nm, respectively, resulting in minimal interference from autofluorescence in most biological samples.

Applications

Use Amplex™ Red Assays for a broad range of investigations, including:

• Cell signaling and lipid studies
• Neurobiology research
• Inflammation and immune function assays
• Metabolism studies

Additional related reagents:

• Amplex™ UltraRed Reagent (Cat. No. A36006): second-generation reagent with greater sensitivity and brightness
• Amplex™ Red/UltraRed Stop Reagent (Cat. No. A33855): enables controlled termination of fluorescence reaction; signal remains stable for at least 3 hours

Custom assay design and packaging options are available.

Specifications

참고 링크

Thermo Fisher Fluorescence Microplate Assays

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