Invitrogen™ Amplex™ Red Hydrogen Peroxide/Peroxidase Assay Kit

Invitrogen™ Amplex™ Red Hydrogen Peroxide/Peroxidase Assay Kit

The Amplex™ Red Hydrogen Peroxide/Peroxidase Assay Kit contains a sensitive, one-step assay that uses the Amplex™ Red reagent (10-acetyl-3,7-dihydroxyphenoxazine) to detect hydrogen peroxide (H2O2) or peroxidase activity. The Amplex™ Red reagent, in combination with horseradish peroxidase (HRP), has been used to detect H2O2 released from biological samples, including cells, or generated in enzyme-coupled reactions. Furthermore, Amplex™ Red reagent can be used as an ultrasensitive assay for peroxidase activity when H2O2 is in excess. In the presence of peroxidase, the Amplex™ Red reagent reacts with H2O2 in a 1:1 stoichiometry to produce the red-fluorescent oxidation product, resorufin. Resorufin has excitation and emission maxima of approximately 571 nm and 585 nm, respectively, and because the extinction coefficient is high (58,000 ± 5,000 cm–1M–1), you can perform the assay fluorometrically or spectrophotometrically. This reaction has been used to detect as little as 10 picomoles of H2O2 in a 100 µL volume or 1 x 10–5 U/mL of HRP

Features

• Sensitive one-step assay to detect hydrogen peroxide or peroxidase in vitro
• Can detect as little as 10 picomoles of hydrogen peroxide or 10 µU/mL of horseradish peroxidase activity in a 100 µL assay volume
• Provides sufficient reagents for approximately 500 assays using reaction volumes of 100 µL per assay
• Designed for a 96-well fluorescence or absorbance microplate reader, but can be easily adapted for other volumes, for fluorimeters, and for spectrophotometer
• For Use With (Equipment): Microplate Reader, Spectrophotometer, Fluorometer