Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Cell culture supernatant
- Sensitivity: 30 nM
Product nameHydrogen Peroxide Assay Kit (Fluorometric - Near Infrared)
See all Hydrogen Peroxide kits
Sample typeCell culture supernatant, Adherent cells
Assay typeEnzyme activity
Species reactivityReacts with: Mammals, Other species
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.
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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 500 tests Assay Buffer 1 x 100ml DMSO 1 x 0.5ml Horseradish Peroxidase 1 x 20 units Hydrogen Peroxide Solution 1 x 200µl IR Peroxidase Indicator 1 vial
RelevanceHydrogen Peroxide (H2O2) is a reactive oxygen metabolic byproduct that serves as a key regulator for a number of oxidative stress-related states. Functioning through NF-kB and other factors, hydroperoxide-mediated pathways have been linked to asthma, inflammatory arthritis, atherosclerosis, diabetic vasculopathy, osteoporosis, neuro-degenerative diseases, Down’s syndrome and immune system diseases.
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.
ab138886 has been referenced in 1 publication.
- Das T et al. Conditions Under Which Glutathione Disrupts the Biofilms and Improves Antibiotic Efficacy of Both ESKAPE and Non-ESKAPE Species. Front Microbiol 10:2000 (2019). PubMed: 31543871