Specifications

  • Assay Type Detection Kit
  • Sample Types Urine, Buffer, Tissue Culture Media
  • Sensitivity 0.038 µM
  • Species All Species
  • Assay Duration 15 Minutes
  • Samples/Plate 40 in duplicate
  • Readout Fluorescent, 590 nm Emission / 520 nm Excitation
  • Standard Curve Hydrogen Peroxide (H2O2) Fluorescent Detection Kit
  • Description

    Assay Principle: 

    The Hydrogen Peroxide (H2O2) Fluorescent Detection Kit quantitatively measures H2O2 levels in urine, buffer, and tissue culture media. The Hydrogen Peroxide (H2O2) Fluorescent Detection Kit is a Detection Kit with a run time of 15 minutes. Please read the complete kit insert for more information before performing this assay.

    Use our provided Hydrogen Peroxide standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a black microtiter plate. Add Fluorescent Detection Reagent and Horseradish Peroxidase Concentrate to each well, tapping the plate to ensure sufficient mixing of reagents. Then, incubate the mixture at room temperature for 15 minutes. The fluorescent reaction occurs between the Fluorescent Detection Reagent, Horseradish Peroxidase Concentrate, and Hydrogen Peroxide within the sample or standard. 

    After the 15-minute incubation, use a plate reader to detect and record the generated fluorescent signal. Use the intensity and the standard curve to calculate the Hydrogen Peroxide concentration in the samples.

    Background:

    In biological systems, incomplete reduction of O2 during respiration produces superoxide anion (O2 -·), which by dismutation superoxide dismutase spontaneously or enzymatically generates H2O2. Many cells produce low levels of O2 -· and H2O2 in response to a variety of extracellular stimuli, such as cytokines (TGF-β1, TNF-α, and various interleukins), peptide growth factors (PDGF, EGF, VEGF, bFGF, and insulin), the agonists of heterotrimeric G protein-coupled receptors (GPCR) such as angiotensin II, thrombin, lysophosphatidic acid, sphingosine 1-phosphate, histamine, and bradykinin, and by shear stress. 

    In 1894, Fenton described the oxidation of tartaric acid by Fe2+ and H2O2. The addition of exogenous H2O2, or the intracellular production in response to receptor stimulation, affects the function of various proteins, including protein kinases, protein phosphatases, transcription factors, phospholipases, ion channels, and G proteins. H2O2 and O2 may participate in the production of singlet oxygen and peroxynitrite. The generation of these species may be concurrent with reactions involving iron, which under some circumstances might be essential contributors to H2O2 toxicity.