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CAT100

Sigma-Aldrich

Catalase Assay Kit

sufficient for ≥100 tests enzymatic, determination of catalase activity in tissues and cells

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84

Quality Level

200

usage

sufficient for ≥100 tests enzymatic

detection method

colorimetric

shipped in

wet ice

storage temp.

2-8°C

Gene Information

human ... CAT(847)

Related Categories

General description

The Catalase Assay Kit contains all necessary components for studying catalase activity in various tissues and subcellular organelles.

Application

Sutitable for Colorimetric and UV Assays

Biochem/physiol Actions

Catalase is a ubiquitous antioxidant enzyme which catalyses the decomposition of hydrogen peroxide (H2O2) to water and oxygen. Hydrogen peroxide is formed in the eukaryotic cell as a by-product of various oxidases and superoxide dismutases. Hydrogen peroxide accumulation in cells causes oxidation of cellular targets such as DNA, proteins, and lipids leading to mutagenesis and cell death. Removal of the H2O2 from the cell by catalase provides protection against oxidative damage to living cells and its role in oxidative stress related diseases has been widely studied.

Features and Benefits

  • Useful for determining catalase activity - may be used in various tissues and cells
  • Simple, optimized protocol - A simple colorimetric assay for analysis of peroxisome enrichment and catalase activity

Suitability

Suitable for studying catalase activity in various tissues and subcellular organelles.

Principle

This assay method is based on the measurement of the hydrogen peroxide substrate remaining after the action of catalase. First, the catalase converts hydrogen peroxide to water and oxygen (catalatic pathway) and then this enzymatic reaction is stopped with sodium azide. An aliquot of the reaction mix is then assayed for the amount of hydrogen peroxide remaining by a colorimetric method.10 The colorimetric method uses a substituted phenol (3,5-dichloro-2-hydroxybenzenesulfonic acid), which couples oxidatively to 4-aminoantipyrine in the presence of hydrogen peroxide and horseradish peroxidase (HRP) to give a red quinoneimine dye (N-(4-antipyryl)-3-chloro-5-sulfonatep-benzoquinone-monoimine) that absorbs at 520 nm

Unit Definition

One unit of catalase will decompose 1.0 micromole of hydrogen peroxide to oxygen and water per minute at pH 7.0 at 25 °C at a substrate concentration of 50 mM hydrogen peroxide.

Preparation Note

Use ultrapure water in preparation of all solutions.

Kit Components Also Available Separately

Product No.
Description
SDS
  • P6782Peroxidase from horseradish, Type VI-A, essentially salt-free, lyophilized powder, ≥250 units/mg solid (using pyrogallol), 950-2000 units/mg solid (using ABTS) 5 mgSDS
  • 323381Hydrogen peroxide solution, contains ~200 ppm acetanilide as stabilizer, 3 wt. % in H2OSDS

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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M B Hampton et al.
FEBS letters, 414(3), 552-556 (1997-10-10)
The induction of apoptosis in Jurkat T-lymphocytes with 50 microM hydrogen peroxide was associated with caspase activation. Caspase activity was first detected 3 h after treatment, and the morphological features of apoptosis were apparent by 6 h. At higher concentrations
M Zámocký et al.
Progress in biophysics and molecular biology, 72(1), 19-66 (1999-08-14)
This review gives an overview about the structural organisation of different evolutionary lines of all enzymes capable of efficient dismutation of hydrogen peroxide. Major potential applications in biotechnology and clinical medicine justify further investigations. According to structural and functional similarities
S Tada-Oikawa et al.
FEBS letters, 442(1), 65-69 (1999-01-29)
Pulsed field gel electrophoresis showed that the initiation time of DNA breakage induced by the DNA alkylating agent duocarmycin A, which is not a redox-cycling agent, was almost the same in the human leukemia cell line HL-60 and its H2O2-resistant
M Ding et al.
Journal of cell science, 113 ( Pt 13), 2409-2419 (2000-06-15)
The intracellular protozoan parasite Toxoplasma gondii, like all members of the phylum Apicomplexa, is known to possess many organelles: in addition to mitochondria and the compartments of the secretory pathway, there is a reduced chloroplast (the apicoplast) and the phylum-specific
A J Kowaltowski et al.
FEBS letters, 473(2), 177-182 (2000-05-17)
The involvement of reactive oxygen species in Ca(2+)-induced mitochondrial membrane permeabilization and cell viability was studied using yeast cells in which the thioredoxin peroxidase (TPx) gene was disrupted and/or catalase was inhibited by 3-amino-1,2, 4-triazole (ATZ) treatment. Wild-type Saccharomyces cerevisiae
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Articles

Organelle Isolation

The isolation of subcellular fractions by centrifugation is a commonly used technique and is widely applicable across multiple cell and tissue types. Because organelles differ in their size, shape, and density, centrifugation can be easily employed to separate and purify organelle fractions from gently homogenized samples.

Mitochondrial Stress and ROS

Oxidative stress is mediated, in part, by reactive oxygen species produced by multiple cellular processes and controlled by cellular antioxidant mechanisms such as enzymatic scavengers or antioxidant modulators. Free radicals, such as reactive oxygen species, cause cellular damage via cellular.

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