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Specifications
- Assay Type Activity Assay
- Sample Types Serum, Plasma, Cells, Tissues, RBC/Erythrocyte Lysates
- Sensitivity 0.073 U/mL
- Species Identical across all species
- Assay Duration 45 Minutes
- Samples/Plate 41 in Duplicate
- Readout Fluorescent, 590 nm emission / 520 nm excitation
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Standard Curve
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Description
Assay Principle:
The Catalase Fluorescent Activity Kit quantitatively measures Catalase activity in serum, plasma, cells, tissues, and RBC/erythrocyte lysates. The Catalase Fluorescent Activity Kit is an Activity Assay with a run time of 45 minutes. Please read the complete kit insert for more information before performing this assay.
Use our provided Catalase Standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate. Add the Hydrogen Peroxide Reagent to each well, tapping the plate to ensure sufficient mixing of reagents. Then incubate the mixture at room temperature for 30 minutes. After the 30-minute incubation, add the Fluorescent Detection Reagent and the HRP Reagent to each well and incubate the plate for 15 minutes. The fluorescent-generating reaction occurs between the HRP, hydrogen peroxide, and the colorless substrate within the sample or standard.
After the 45-minute incubation, use a plate reader to detect and record the generated signal at 590nm. Use the intensity and the standard curve to calculate the catalase activity in the samples.
Background:
Hydrogen peroxide (H2O2) is one of the most frequently occurring reactive oxygen species. It is formed either in the environment, as a by-product of aerobic metabolism, superoxide formation, and dismutation, or as a product of oxidase activity. Both excessive hydrogen peroxide and its decomposition product, hydroxyl radical, are harmful to most cell components. Its rapid removal is essential for all aerobically living prokaryotic and eukaryotic cells. One of the most efficient ways of removing peroxide is through the enzyme catalase, which is encoded by a single gene and is highly conserved among species. Mammals, including humans and mice, express catalase in all tissues. A high catalase concentration exists in the liver, kidneys, and erythrocytes. Transcription, post-transcription, and post-translation regulation control catalase expression. Peroxisomes, cancers, and thyroid dysfunction all exhibit high catalase activity.