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, H2
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 H2
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.