Overview

  • Product name

    Hydrogen Peroxide Assay Kit (Cell-based)
    See all Hydrogen Peroxide kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Sensitivity

    3 µM
  • Assay time

    0h 30m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Hydrogen Peroxide Assay Kit (Cell-based) (ab138874) uses our AbGreen Indicator to quantify hydrogen peroxide in live cells. The indicator is cell-permeable, and generates green fluorescence when it reacts with hydrogen peroxide. 


    The assay is in a “mix and read” assay format that is compatible with HTS liquid handling instruments and a 96-well or 384-well plate format. It provides a sensitive, one-step fluorometric assay to detect as little as 0.3 nanomoles of H2O2 in a 100 µL assay volume (3 µM). Results can be read by either a fluorescence microplate reader at Ex/Em = 490/520 nm for H2O2 detection in solution or a fluorescence microscopy for live cell H2O2 imaging.


    Hydrogen peroxide assay protocol summary:
    - add samples and standards to wells
    - add green indicator solution
    - analyze with microplate reader for 15-60 min.

    Alternatively, for live cell assay:
    - add green indicator solution to cells
    - incubate for 5-60 min whilst analyzing with microplate reader or fluorescence microscope

  • 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 many biological events that are linked to asthma, atherosclerosis, diabetic vasculopathy, osteoporosis, a number of neurodegenerative diseases and Down’s syndrome. The measurement of this reactive species is helpful for determining how oxidative stress modulates various intracellular pathways.

  • Platform

    Fluorescence microscope

Properties

Images

  • Fluorescence images of intracellular hydrogen peroxide in HeLa cells using Hydrogen Peroxide Assay Kit (Cell-based) (ab138874). HeLa cells were treated with (left) or without (right) 100 µM hydrogen peroxide at 37ºC for 90 minutes.

Protocols

References

This product has been referenced in:

  • Zhou T  et al. Growth suppression and apoptosis-like cell death in Microcystis aeruginosa by H2O2: A new insight into extracellular and intracellular damage pathways. Chemosphere 211:1098-1108 (2018). Read more (PubMed: 30223325) »
See 1 Publication for this product

Customer reviews and Q&As

User Friendly Kit With Good Results

Good Excellent 5/5 (Ease of Use)
Abreviews
Our lab ordered this kit to be used in RAW 264.7 (murine macrophage) cells that were treated with different metals at different time points. The kit is predicted to work with mammalian species, so yes, it will work for the RAW 264.7 cell line. The directions are very easy to follow, and we were able to detect as little as 3 uM of Hydrogen Peroxide production in live cells. We will definitely be ordering this kit again since we did not take images this time around.

Dr. Nicole Olgun

Verified customer

Submitted Oct 01 2015

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