Overview

  • Product name

    Hydrogen Peroxide Assay Kit (Fluorometric - Near Infrared)
    See all Hydrogen Peroxide kits
  • Detection method

    Fluorescent
  • Sample type

    Cell culture extracts
  • Assay type

    Enzyme activity
  • Sensitivity

    30 nM
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Hydrogen Peroxide Assay Kit (Fluorometric - Near Infrared) (ab138886) uses our unique AbIR Peroxidase Indicator substrate to quantify hydrogen peroxide in solutions and cell extracts. AbIR Peroxidase Indicator generates the fluorescence that is pH-independent from pH 4 to 10. It is a superior alternative to ADHP for the detections that require low pH where ADHP has reduced fluorescence. In addition, AbIR Peroxidase Indicator generates a product that has maximum absorption at 647 nm with maximum emission at 670 nm. This near infrared fluorescence minimizes the assay background that is often caused by the autofluorescence of biological samples. It can also be used to detect a variety of oxidase activities through enzyme-coupled reactions.


    This is an optimized “mix and read” assay that is compatible with HTS liquid handling instruments. It provides a sensitive, one-step fluorometric assay to detect as little as 3 pmol H2O2 /100 µL assay volume (30 nM). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. Its signal can be easily read by either a fluorescence microplate reader at Ex/Em = ~640/680 nm or an absorbance microplate reader at ~650 nm. Due to its long emission wavelength, this kit has low interference from biological samples.

  • Notes

    Hydrogen peroxide (H2O2) is a reactive oxygen metabolic by-product that serves as a key regulator for a number of oxidative stress-related states. It is involved in a number of biological events that have been linked to asthma, atherosclerosis, diabetic vasculopathy, osteoporosis, a number of neurodegenerative diseases and Down's syndrome. Perhaps the most intriguing aspect of H2O2 biology is the recent report that antibodies have the capacity to convert molecular oxygen into hydrogen peroxide to contribute to the normal recognition and destruction processes of the immune system. Measurement of this reactive species will help to determine how oxidative stress modulates a variety of intracellular pathways.

  • Platform

    Microplate reader

Properties

Images

  • Hydrogen Peroxidase dose response was measured on a 96-well black plate with Hydrogen Peroxidase Assay Kit (Fluorometric-Near Infrared), using a fluorescence microplate reader (Molecular Devices). As low as 0.03 µM H2O2 can be detected with 1 minute incubation time (n=3). The insert shows the low levels of Hydrogen Peroxidase detection.

Protocols

References

ab138886 has not yet been referenced specifically in any publications.

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