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Catalase colorimetric activity kit

Manufactured by Thermo Fisher Scientific
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The Catalase Colorimetric Activity Kit is a laboratory product designed to measure the activity of the enzyme catalase. Catalase is an important enzyme found in most living organisms that functions to catalyze the decomposition of hydrogen peroxide into water and oxygen. The kit provides a colorimetric assay method to quantify catalase activity in a variety of sample types.

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Lab products found in correlation

Protein assay kit, by Bio-Rad (2 mentions) Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions) Tac colorimetric assay kit, by Abcam (1 mentions) Superoxide dismutase sod colorimetric activity kit, by Thermo Fisher Scientific (1 mentions) Lipid peroxidation mda assay kit, by Merck Group (1 mentions) Dna damage 8 ohdg elisa kit, by StressMarq Biosciences (1 mentions)

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Catalase (10 citations) Colorimetric activity kit (5 citations)

13 protocols using catalase colorimetric activity kit

1

Cisplatin-induced Catalase Activity

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RPTEC/hTERT1 cells were cultured in 6-well plates (250,000 cells/well) and exposed for 48 hours to cisplatin (50 μM) with or without KW6002 (25 μM). Catalase activity was assessed using the Catalase Colorimetric Activity Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions.
Dewaeles E., Carvalho K., Fellah S., Sim J., Boukrout N., Caillierez R., Ramakrishnan H., Van der Hauwaert C., Vijaya Shankara J., Martin N., Massri N., Launay A., Folger J.K., de Schutter C., Larrue R., Loison I., Goujon M., Jung M., Le Gras S., Gomez-Murcia V., Faivre E., Lemaire J., Garat A., Beauval N., Maboudou P., Gnemmi V., Gibier J.B., Buée L., Abbadie C., Glowacki F., Pottier N., Perrais M., Cunha R.A., Annicotte J.S., Laumet G., Blum D, & Cauffiez C. (2022). Istradefylline protects from cisplatin-induced nephrotoxicity and peripheral neuropathy while preserving cisplatin antitumor effects. The Journal of Clinical Investigation, 132(22), e152924.
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2

Catalase Activity Assay in E. coli on GaN

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The catalase colorimetric activity kit from Thermo Fisher (catalog number: EIACATC) was utilized according to the supplied protocol. A catalase standard was provided to generate a standard curve. The E. coli cultures were grown overnight and diluted to an OD600 of 0.1. Subsequently, 100 μL aliquots were added to the UV treated and nontreated GaN surfaces. The cells were then vortexed loose from the surface of the GaN and diluted in the assay buffer, 25 μL of which was added to the wells of a 96-well plate. 25 μL of hydrogen peroxide (H2O2) was then added to each well and left to incubate at room temperature for 30 min. 25 μL of the provided substrate and horseradish peroxidase (HRP) were then added and left to incubate further for 15 min. The HRP and substrate reacted to form a pink solution. The 96-well plate was then read spectrophotometrically at OD560. An increase in catalase activity is seen by the decrease in pinkness of the samples, which indicates a decrease in the amount of H2O2. The number of units of catalase present in each sample was calculated using the initially generated standard curve. Each unit of catalase corresponds to the decomposition of 1 μmol of H2O2 per minute at room temperature.
Iyer D., Gulyuk A.V., Reddy P., Kirste R., Collazo R., LaJeunesse D.R, & Ivanisevic A. (2019). Behavior of E. coli with Variable Surface Morphology Changes on Charged Semiconductor Interfaces. ACS applied bio materials, 2(9), 4044-4051.
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3

Catalase Activity Quantification in Hemolymph

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Catalase activity in hemolymph samples was determined using a catalase colorimetric activity kit (Invitrogen, Frederick, MD, USA) according to the manufacturer’s instructions. Catalase concentration (U/mL) at 560 nm was measured (n = 3) using an EpochTM Microplate Spectrophotometer (Epoch 2, BioTek, Winooski, VT, USA).
Hossen S., Sukhan Z.P., Kim S.C., Hanif M.A., Kong I.K, & Kho K.H. (2023). Molecular Cloning and Functional Characterization of Catalase in Stress Physiology, Innate Immunity, Testicular Development, Metamorphosis, and Cryopreserved Sperm of Pacific Abalone. Antioxidants, 12(1), 109.
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4

Quantification of Antioxidant Enzymes and Capacity

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The enzyme activity of superoxide dismutase (SOD) and catalase (CAT) were measured quantitatively using their respective kits from Invitrogen™ (Waltham, MA, USA): superoxide dismutase (SOD) colorimetric activity kit and catalase colorimetric activity kit. Following the manufacturer’s protocol, the activity (U/mL) of antioxidant enzymes was measured in 150 µg of protein samples at 450 nm and 560 nm using standard curves for SOD and CAT, respectively. The total antioxidant capacity (TAC) colorimetric assay kit (BioVision, Milpitas, CA, USA) was used to detect small antioxidant molecules in the presence of a proprietary protein mask. Following the manufacturer’s protocol, the levels of small antioxidants were measured in a 150-µg protein sample using a 96-well plate. The absorbance was plotted at 570 nm as a function of Trolox concentration (nmol).
Almarzouq D, & Al-Maghrebi M. (2023). NADPH Oxidase-Mediated Testicular Oxidative Imbalance Regulates the TXNIP/NLRP3 Inflammasome Axis Activation after Ischemia Reperfusion Injury. Antioxidants, 12(1), 145.
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5

Quantifying Brain Catalase Activity

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Catalase activity was determined in the brain regions using the Catalase Colorimetric Activity Kit (Invitrogen™, Waltham, MA, USA). A bovine catalase standard was used to generate a standard curve for the assay and all tissue lysates read off the standard curve. Brain lysates were diluted at 1:20 in the provided assay buffer and added to the wells of a half-area transparent plate. H2O2 was added to each well, and the plate was incubated at room temperature for 30 min. Following this incubation, the supplied colorimetric detection reagent was added, followed by diluted horseradish peroxidase and incubation at room temperature for 15 min. The colored product was read at 560 nm. All brain extracts were assayed in triplicate, with the CV of replicate measures being <10%. The activity of catalase was standardized to protein and expressed as U/mg protein.
Kanon A.P., Giezenaar C., Roy N.C., Jayawardana I.A., Lomiwes D., Montoya C.A., McNabb W.C, & Henare S.J. (2024). Effects of Green and Gold Kiwifruit Varieties on Antioxidant Neuroprotective Potential in Pigs as a Model for Human Adults. Nutrients, 16(8), 1097.
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6

Quantification of Catalase Activity in WAT

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The total catalase activity was measured in the WAT using the Catalase Colorimetric Activity Kit (Invitrogen, EIACATC; Waltham, MA, USA), according to the manufacturer’s protocol. Briefly, the WAT was homogenised in 0.5 mL cold 1× Assay Buffer per 100 mg of tissue using the TissueLyser II. The lysates were centrifuged at 10,000× g for 15 min at 4 °C. Avoiding the lipid layer, the cleared lysates were transferred to fresh tubes. The total protein content was quantitated using a DC protein assay, and the samples were normalised to 1.6 μg/mL with 1× Assay Buffer. The assay was then carried out according to the kit protocol. GraphPad Prism 9 software was used to generate the standard curve and interpolate the catalase activity values for the samples.
Croft A.J., Kelly C., Chen D., Haw T.J., Sverdlov A.L, & Ngo D.T. (2023). Overexpression of Mitochondrial Catalase within Adipose Tissue Does Not Confer Systemic Metabolic Protection against Diet-Induced Obesity. Antioxidants, 12(5), 1137.
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7

Quantifying Catalase Activity in Cells

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Single-cell suspensions were prepared from spleen and lung, and CD11c positive/negative cells were isolated as needed using CD11c ultrapure magnetic beads as described above. Cells were lysed by quick freeze/thaw followed by sonication using an Ultrasonic Liquid Processor (Misonix Inc, model XL2020) under the setting of 30% amplitude/30s on/30s off over 4 cycles. The resulting homogenate was centrifuged at 10,000 ×g for 15 min at 4°C. The supernatants were collected to measure catalase activity using a Catalase Colorimetric Activity Kit (Invitrogen, catalog EIACATC). Protein concentration in each sample was determined using a Protein Assay kit (BioRad). Catalase activity was normalized to protein concentration (U/mg).
Yuan H., Chen J., Hu S., Oriss T.B., Kale S.L., Das S., Nouraie S.M., Ray P, & Ray A. (2021). Early life exposure to house dust mite allergen prevents experimental allergic asthma requiring mitochondrial H2O2. Mucosal immunology, 15(1), 154-164.
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8

Serum Catalase Activity Assay

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Activity of serum catalase was measured by Catalase colorimetric activity kit (EIACATC, Invitrogen). Serum and standard were prepared according to the kit. 25 µl of standard and serum sample was added to wells in a microtitre plate followed by addition of hydrogen peroxide. After incubating for 30 min, substrate and HRP added and read at 560 nm. SOD activity was expressed as U/ml.
Abudawood M., Tabassum H., Almaarik B, & Aljohi A. (2019). Interrelationship between oxidative stress, DNA damage and cancer risk in diabetes (Type 2) in Riyadh, KSA. Saudi Journal of Biological Sciences, 27(1), 177-183.
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9

Catalase Activity Measurement in RAW Cells

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Catalase (CAT) activity was evaluated using the catalase colorimetric activity kit (Invitrogen Co.). RAW 264.7 cells were seeded in a 6-well plate at a concentration of 2 × 105 cells/well. After incubation at 37 °C with 5% CO2 for 4 h, the cells were treated with AHL and AHR extracts at various concentrations (100, 250, and 500 μg/mL) and were cultured in the incubator for 48 h. Cells were centrifuged at 1500 rpm at 4 °C for 10 min and the supernatant was transferred to a new plate. Briefly, 25 μL of samples were mixed with 25 μL of hydrogen peroxide reagent and were reacted at RT for 30 min. The reaction product was added to 25 μL of substrate solution and HRP solution and was incubated at 25 °C for 15 min. The absorbance was measured at 560 nm using a microplate reader (Molecular Devices). CAT activity was calculated by the standard curve.
Jeong U.Y., Jung J., Lee E.B., Choi J.H., Kim J.S., Jang H.H., Park S.Y, & Lee S.H. (2022). Antioxidant and Immune Stimulating Effects of Allium hookeri Extracts in the RAW 264.7 Cells and Immune-Depressed C57BL/6 Mice. Antioxidants, 11(10), 1927.
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10

Catalase Activity Assay Protocol

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The Catalase activity was assayed using the Catalase colorimetric activity kit from Invitrogen (Life Technologies, Carlsbad, CA, USA). Briefly the platelet lysate was diluted 1:10 in the Assay Buffer provided and assayed as per the manufacturer's instructions. One unit of catalase (U) decomposes 1.0 μmol of H2O2 per min at pH 7.0 and 25 °C. The results are expressed as U/mg protein.
Jain K., Tyagi T., Patell K., Xie Y., Kadado A.J., Lee S.H., Yarovinsky T., Du J., Hwang J., Martin K.A., Testani J., Ionescu C.N, & Hwa J. (2019). Age associated non-linear regulation of redox homeostasis in the anucleate platelet: Implications for CVD risk patients. EBioMedicine, 44, 28-40.
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