EnzyChrom™ Catalase Assay Kit

Protocol SDS

Application

• For quantitative determination of catalase activity and evaluation of drug effects on catalase activity.

Key Features

• Sensitive and accurate. Use 10 µL sample. Linear detection range 0.2 to 5 U/L catalase activity.
• Simple and Convenient. The procedure involves adding a Substrate to the sample, incubation for 30 min, followed by a Detection Reagent and reading the optical density or fluorescence intensity.

Method

• OD570nm, or FL530/585nm

Samples

• Serum, plasma, urine, saliva, cell culture etc

Species

• All

Procedure

• 40 min

Size

• 100 tests

Detection Limit

• 0.2 U/L

Shelf Life

• 6 months

More Details

CATALASE (EC 1.11.1.6), is an ubiquitous antioxidant enzyme that catalyzes the decomposition of hydrogen peroxide (H₂O₂) to water and oxygen. By preventing excessive H₂O₂ build-up, catalase allows important cellular processes which produce H₂O₂ as a byproduct to occur safely. Deficiency in catalase activity has been associated with grey hair and peroxisomal disorder acatalasia. Simple, direct and high-throughput assays for catalase activity find wide applications. BioAssay Systems improved assay directly measures catalase degradation of H₂O₂ using a redox dye. The change in colour intensity at 570nm or fluorescence intensity (λex/em = 530/585nm) is directly proportional to the catalase activity in the sample.

Regarding the technique used in the DPOD-100 peroxidase assay and the ECAT-100 catalase assay that seems to be really similar, how will we be sure that one of them will measure the catalase activity and the other the peroxidase?

The DPOD assay measures the conversion of the dye reagent and hydrogen peroxide, which are provided in large excess as substrates for cellular peroxidases. Endogenous catalase activity does not interfere with the assay, because it does not react with the dye reagent, hydrogen peroxide is in large excess, and the reaction is short (10 minutes), leaving the H₂O₂ concentration practically unchanged.

The ECAT assays measures catalase activity by first incubating much lower concentrations of hydrogen peroxide with the samples for 30 minutes. Then the decrease in hydrogen peroxide is measured by adding the dye reagent and HRP in excess.

You will have 100 µL of each standard when you have pipetted all solutions for the standard curve. How stable are the 100 µl standards? Can they be used the next day to make another standard curve or can the values from the already made standard curve be used when you have other samples the next day?

The H₂O₂ standards are stable for an hour. Therefore assays should be run as soon as possible with the samples and standards. It is recommended that customers run standard curve in each experiment.

AlAsmari, A. F.,et al (2020). Elucidation of the molecular mechanisms underlying sorafenib-induced Hepatotoxicity. Oxidative Medicine and Cellular Longevity, 2020, 1-10. Assay: Catalase in rat liver tissue.

Paul, S.,et al. (2021). D4F prophylaxis enables redox and energy homeostasis while preventing inflammation during hypoxia exposure. Biomedicine & Pharmacotherapy, 133, 111083. Assay: Catalase in rat tissue.

Humam, A. M., et al. (2021). Supplementation of postbiotic RI11 improves antioxidant enzyme activity, upregulated gut barrier genes, and reduced cytokine, acute phase protein, and heat shock protein 70 gene expression levels in heat-stressed broilers. Poultry Science, 100(3), 100908. Assay: Catalase in chicken plasma.

AlAsmari, A. F., et al (2020). Liraglutide attenuates gefitinib-induced cardiotoxicity and promotes cardioprotection through the regulation of MAPK/NF-kb signaling pathways. Saudi Pharmaceutical Journal, 28(4), 509-518. Assay: Catalase in rat heart tissue.

Wahyuningsih, S. P., et al. (2020). Antioxidant and Nephroprotective effects of okra pods extract (Abelmoschus esculentus L.) against lead acetate-induced toxicity in mice. Scientifica, 2020, 1-10. Assay: Catalase in mouse serum.

Sahin, T. D., et al. (2020). Infliximab prevents dysfunction of the vas deferens by suppressing inflammation and oxidative stress in rats with chronic stress. Life Sciences, 250, 117545. Assay: Catalase in rat tissue.

Jain, S., et al (2020). Saliva panel of protein candidates: A comprehensive study for assessing high altitude acclimatization. Nitric Oxide, 95, 1-11. Assay: Catalase in human saliva.

Nawab, A.,et al. (2019). Effect of dietary curcumin on the antioxidant status of laying hens under high- temperature condition. Journal of Thermal Biology, 86, 102449. Assay: Catalase in chicken serum.

Xin, X., et al. (2020). Exploring the biophysicochemical alteration of green alga Asterococcus superbus interactively affected by nanoparticles, triclosan and illumination. Journal of Hazardous Materials, 398, 122855. Assay: Catalase in algae cells.

Pena-Bermudez, Y. A.,et al (2020). Effects of feeding increasing levels of yerba mate on lamb meat quality and antioxidant activity. Animals, 10(9), 1458. Assay: Catalase in sheep tissue.

Pathirana, R. U., et al (2020). Longer ubiquinone side chains contribute to enhanced farnesol resistance in yeasts. Microorganisms, 8(11), 1641 Assay: Catalase in yeast cells.

Lin, Ting-An, et al (2019). Red Quinoa Bran Extracts Protects against Carbon Tetrachloride-Induced Liver Injury and Fibrosis in Mice via Activation of Antioxidative Enzyme Systems and Blocking TGF-beta1 Pathway. Nutrients 11.2: 395. Assay: Catalase in mice liver tissue.

Cueno, Marni E., and Kuniyasu Ochiai (2018). Gingival periodontal disease (PD) level-butyric acid affects the systemic blood and brain organ: insights into the systemic inflammation of periodontal disease. Frontiers in immunology 9:1158. Assay: Catalase in rat blood cytosol.

Kang, Changwoo, et al (2018). Therapeutic effect of ascorbic acid on dapsone-induced methemoglobinemia in rats. Clinical and experimental emergency medicine 5.3: 192. Assay: Catalase in rat plasma.

Salvatori, Ornella, et al (2018). Candida albicans Ras1 inactivation increases resistance to phagosomal killing by human neutrophils. Infection and immunity 86.12: e00685-18. Assay: Catalase in Candida albicans cells.

Camini, Fernanda Caetano, et al (2017). Oxidative stress in Mayaro virus infection. Virus research 236: 1-8. Assay: Catalase in human cells.

Kang, Hyeon Hui, et al (2017). Chronic intermittent hypoxia induces liver fibrosis in mice with diet-induced obesity via TLR4/MyD88/MAPK/NF-kB signaling pathways. Biochemical and biophysical research communications 490.2: 349-355. Assay: Catalase in mice tissues.

Kook, Sung-Ho, et al (2017). Dietary hydroxycinnamates prevent oxidative damages to liver, spleen, and bone marrow cells in irradiation-exposed mice. Food science and biotechnology 26.1: 279-285. Assay: Catalase in mice tissues.

Oliveira, C. S., et al (2017). Moderate aerobic exercise on the recovery phase of gentamicin-induced acute kidney injury in rats. Life sciences 169: 37-42. Assay: Catalase in rat tissues.

Pradhan, Arnab, et al (2017). Elevated catalase expression in a fungal pathogen is a double-edged sword of iron. PLoS pathogens 13.5: e1006405. Assay: Catalase in C. albicans cells.

Cueno ME (2013). Orally supplemented catechin increases heme amounts and catalase activities in rat heart blood mitochondria: a comparison between middle-aged and young rats. Exp Gerontol. 48(11):1319-22. Assay: Catalase in rat blood mitochondria.

Chiu CC et al (2012). Beneficial Effects of Ocimum gratissimum Aqueous Extract on rats with CCl(4)-Induced Acute Liver Injury. Evid Based Complement Alternat Med 2012:736752. Assay: Catalase in rat serum.

Hadzi-Petrushev N et al (2012). L-2-oxothiazolidine-4-carboxylate influence on age- and heat exposure-dependent peroxidation in rat’s liver and kidney. J Therm Biol 37(5):361-365. Assay: Catalase in rat Kidney.

Hadzi-Petrushev N et al (2012). L-2-oxothiazolidine-4-carboxylate influence on age- and heat exposure-dependent peroxidation in rat’s liver and kidney. J Therm Biol 37(5):361-365. Assay: Catalase in rat Kidney.

Zhu T et al (2012) Effects of the iron-chelating agent deferoxamine on triethylene glycol dimethacrylate, 2-hydroxylethyl methacrylate, hydrogen peroxide-induced cytotoxicity. J Biomed Mater Res B Appl Biomater 100(1):197-205. Assay: Catalase in human dental pulp cells.

Hadzi-Petrushev N et al (2011) L-2-oxothiazolidine-4-carboxylate influence on age- and heat exposure-dependent redox changes in rat’s blood plasma. J Physiol Sci 61(5):437-442. Assay: Catalase in rat plasma.

Labib, HM, et al (2010). The Role of Oxidative Stress Markers and Nitric Oxide Levels in the Pathogenesis of Glaucoma. Austr. J. Basic and Applied Sci 4(8): 3553-3558. Assay: Catalase in human blood.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.