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Hydrogen peroxide assay
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 and intracellular pathways that have been linked to several diseases. Measurements of reactive oxygen species help to determine how oxidative stress modulates varied intracellular pathways.
Chemediate’s Kit uses a non-fluorescent peroxidase substrate to quantify hydrogen peroxide in solution, in cell extracts and released from certain live cells, Catalyzed by peroxidase, H2O2 oxidizes 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) in a 1:1 stoichiometry to produce a fluorescent product resorufin.  The one-step fluorometric Hydrogen Peroxide Assay can detect as low as 30nM of H2O2 released from cells or enzyme coupled reactions.  The assay can be performed in a convenient 96-well or 384-well microplate format and is amenable to either fluorescence or absorbance based detection platforms with Ex/Em = 560 nm/585 nm or absorbance at 570 nm. 

Figure 1: Hydrogen peroxide titration curve using ADHP as the co-substrate of horse-radish peroxidase, as low as 30 nM of H2O2 can be detected with 15 minutes incubation.
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