Amplite® Colorimetric Hydrogen Peroxide Assay Kit

Amplite® Colorimetric Hydrogen Peroxide Assay Kit is a colorimetric assay kit for quantifying hydrogen peroxide in solutions and cell extracts using a near-infrared absorbance readout.

Colorimetric detection: Uses Amplite® IR peroxidase substrate to generate an intense blue product in the presence of hydrogen peroxide.
Low-background readout: Near-infrared absorbance minimizes interference from biological samples.
Broad pH compatibility: Produces a pH-independent signal from pH 4 to 10.
HTS compatible: Mix-and-read workflow is compatible with liquid handling systems and absorbance measurement at 650 nm.
Applications: Suitable for hydrogen peroxide quantitation and oxidase activity measurements through enzyme-coupled reactions.

Hydrogen Peroxide dose response was measured in a a white wall/clear bottom 96-well plate with the Amplite® Colorimetric Hydrogen Peroxide Assay Kit using a Spectramax absorbance microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
11500	500 Tests	Price

Overview

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 varied intracellular pathways. This Amplite® Colorimetric Hydrogen Peroxide Assay Kit uses our unique Amplite® IR peroxidase substrate to quantify hydrogen peroxide in solutions and cell extracts. Upon hydrogen peroxide oxidation the colorless Amplite® IR generates an intense blue color product that is pH-independent from pH 4 to 10. The existing colorimetric hydrogen peroxide assays (from other vendors) often have severe sample interferences caused by the inherent absorption of biological samples. The near infrared absorption of Amplite® IR product minimizes the assay background that since the biological samples rarely absorb light beyond 600 nm. It can also be used to detect a variety of oxidase activities through enzyme-coupled reactions. The kit is an optimized 'mix and read' assay that is compatible with HTS liquid handling instruments.

Citations

View all 8 citations: Citation Explorer

Antioxidant Properties of a Rhodiola rosea Constituent, Rosavin

Authors:

Brink, Drew F and Sapp, Tiara L and Ghafoor, Trwska S and Boyland, Paige A and Tamazawa, Yukako C and Kaur, Guneet and Shults, Nataliia V and Sullivan, Robert D and Suzuki, Yuichiro J

Journal:

Life (2026): 1510

Antioxidant-Thermoresponsive Collagen/Chitosan Nanocomposite Hydrogels With Praseodymium-Doped Mesoporous Bioglass for Enhanced Bone Regeneration

Authors:

Kaur, Kulwinder and Xu, Zhiyan and Boccaccini, Aldo R and Murphy, Ciara M

Journal:

Journal of Biomedical Materials Research Part A (2026): e70128

Anti-inflammatory effects of cold atmospheric plasma irradiation on the THP-1 human acute monocytic leukemia cell line

Authors:

Hirasawa, Ito and Odagiri, Haruka and Park, Giri and Sanghavi, Rutvi and Oshita, Takaya and Togi, Akiko and Yoshikawa, Katsunori and Mizutani, Koji and Takeuchi, Yasuo and Kobayashi, Hiroaki and others,

Journal:

PLOS ONE (2023): e0292267

Macrophage metabolism reprogramming EGCG-Cu coordination capsules delivered in polyzwitterionic hydrogel for burn wound healing and regeneration

Authors:

Li, Qinghua and Song, Huijuan and Li, Shuangyang and Hu, Pengbo and Zhang, Chuangnian and Zhang, Ju and Feng, Zujian and Kong, Deling and Wang, Weiwei and Huang, Pingsheng

Journal:

Bioactive Materials (2023): 251--264

One-dimensional array of nanoparticles using segmented polyurethane nanofibers as a template for enhanced catalytic efficiency

Authors:

Meboso, Taichi and Goto, Satoshi and Takamura, Eiichiro and Sakamoto, Hiroaki

Journal:

Colloid and Polymer Science (2023): 1--7

References

View all 152 references: Citation Explorer

Effect of antisense oligonucleotide against Smac/DIABLO on inhibition of hydrogen peroxide induced myocardial apoptosis of neonatal rats

Authors:

Liang PF, Huang XY, Long JH, Xiao MZ, Yang XH, Zhang PH.

Journal:

Zhonghua Shao Shang Za Zhi (2006): 175

Cardioprotective role of endogenous hydrogen peroxide during ischemia-reperfusion injury in canine coronary microcirculation in vivo

Authors:

Yada T, Shimokawa H, Hiramatsu O, Haruna Y, Morita Y, Kashihara N, Shinozaki Y, Mori H, Goto M, Ogasawara Y, Kajiya F.

Journal:

Am J Physiol Heart Circ Physiol (2006): H1138

Simple and rapid determination of hydrogen peroxide using phosphine-based fluorescent reagents with sodium tungstate dihydrate

Authors:

Onoda M, Uchiyama T, Mawatari K, Kaneko K, Nakagomi K.

Journal:

Anal Sci (2006): 815

Specific aquaporins facilitate the diffusion of hydrogen peroxide across membranes

Authors:

Bienert GP, Moller AL, Kristiansen KA, Schulz A, Moller IM, Schjoerring JK, Jahn TP.

Journal:

J Biol Chem. (2006)

Enzymatic oxidation of dipyridamole in homogeneous and micellar solutions in the horseradish peroxidase-hydrogen peroxide system

Authors:

Almeida LE, Imasato H, Tabak M.

Journal:

Biochim Biophys Acta (2006): 216

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	650 nm
Recommended plate	Clear bottom

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Email	sales@aatbio.com
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Page updated on October 2, 2026

Component A: Amplite® IR Peroxidase Substrate	1 vial
Component B: H2O2	1 vial (3% stabilized solution, 200 μL)
Component C: Assay Buffer	1 bottle (100 mL)
Component D: Horseradish Peroxidase	1 vial (20 units)
Component E: DMSO	1 vial (0.5 mL)