Catalase Assay Kit

N° du produit ABIN1000296

Détail du produit

Antigène: Catalase (CAT)

Application: Activity Assay (AcA)

Type d'échantillon: Serum, Plasma, Urine, Saliva, Cell Culture Supernatant

Specificité: 0.2 U/L

Attributs du produit: 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 sample, incubation for 30 min, followed by a Detection Reagent and reading the optical density or fluorescence intensity.

Ingrédients: Assay Buffer: 25 mL. HRP Enzyme: 120 µL. Dye Reagent: 120 µL. H2O2 Solution: 100 µL. 3% H2O2 Positive Control: 8 µL Catalase.

Matériel non inclus: Pipetting devices, centrifuge tubes, uncoated 96-well plates, optical density plate reader, fluorescence plate reader, homogenizer etc.

Information d'application

Indications d'application: Direct Assays: catalase activity in biological samples e.g. serum, plasma, urine, saliva, cell culture etc.
Drug Discovery/Pharmacology: effects of drugs on catalase activity.

Protocole: 1. Enzyme Reaction: Mix 5 µL 3% H2O2 and 914 µL dH2O (final 4.8 mM). Prepare enough 50 µMH2O2 Substrate for sample, positive control and sample blank by mixing, for each well, 1 µL of the 4.8 mM H2O2 with 95 µL Assay Buffer. Note: diluted H2O2 is not stable. Prepare fresh dilutions for each experiment. Add 90 µL of the 50 µMSubstrate to these wells to initiate the catalase reaction. Tap plate quick to mix. Incubate 30 min at room temperature. During the incubation time, proceed with Steps 3 and 4 below.
2. H2O2 Standard Curve. Mix 40µL of the 4.8 mM H2O2 with 440 µL dH2O to yield 400 µMH2O2. Prepare standards. Transfer 10 µL standards into separate wells of the 96-well plate. Add 90 µL Assay Buffer to the standards.
3. Detection. Prepare enough Detection Reagent by mixing, for each reaction well (Sample, Control and Standard wells), 102 µL Assay Buffer, 1 µL Dye Reagent and 1 µL HRP Enzyme. At the end of the 30 min incubation (Step 2), add 100 µL Detection Reagent per well. Tap plate to mix. Incubate for 10 min.
5. Read optical density at 570nm (550 to 585nm) or fluorescence intensity at em/ex = 585/530nm.

Préparation des réactifs:

Equilibrate all components to room temperature. Briefly centrifuge all tubes before opening. Keep thawed HRP Enzyme on ice. For colorimetric assays, use a clear flat-bottom 96-well plate. For fluorimetric assays, use a solid black flat-bottom 96-well plate. Samples and Controls: transfer 10 µL sample into wells of the 96-well plate. In addition, for each assay run, prepare one sample blank well that contains only 10 µL Assay Buffer. Add 400 µL Assay Buffer to Positive Control tube and mix well. Transfer 10 µL of the reconstituted Positive Control into separate wells. Note: (1). For unknown samples, perform several dilutions to ensure that catalase activity is within the linear range 0.2 to 5 U/L. (2) The provided catalase serves as a positive control to ensure assay is working and should not be used to calculate the Sample catalase activity.

Calcul des résultats:

Subtract blank value (#4) from the standard values and plot the OD or F against standard concentrations. Unit definition: one unit is the amount of catalase that decomposes 1 µMole of H2O2 per min at pH 7.0 and room temperature.

Restrictions: For Research Use only

Stockage

Stock: -20 °C

Référence

Cueno, Tamura, Imai, Ochiai: "Orally supplemented catechin increases heme amounts and catalase activities in rat heart blood mitochondria: a comparison between middle-aged and young rats." dans: Experimental gerontology, Vol. 48, Issue 11, pp. 1319-22, (2013) (PubMed).

Chiu, Huang, Chen, Kao, Liu, Wang, Tzang, Hsu: "Beneficial Effects of Ocimum gratissimum Aqueous Extract on Rats with CCl(4)-Induced Acute Liver Injury." dans: Evidence-based complementary and alternative medicine : eCAM, Vol. 2012, pp. 736752, (2012) (PubMed).

Hadzi-Petrushev, Jankulovski, Hristov, Mladenov: "L-2-oxothiazolidine-4-carboxylate influence on age- and heat exposure-dependent redox changes in rat's blood plasma." dans: The journal of physiological sciences : JPS, Vol. 61, Issue 5, pp. 437-42, (2011) (PubMed).

Zhu, Lim, Park, Son, Yang: "Effects of the iron-chelating agent deferoxamine on triethylene glycol dimethacrylate, 2-hydroxylethyl methacrylate, hydrogen peroxide-induced cytotoxicity." dans: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 100, Issue 1, pp. 197-205, (2011) (PubMed).

Habib, Eisa, Arafat, Marie: "Pulmonary involvement in early rheumatoid arthritis patients." dans: Clinical rheumatology, Vol. 30, Issue 2, pp. 217-21, (2011) (PubMed).

Détail du antigène

Antigène: Catalase (CAT)

Autre désignation: Catalase (CAT Produits)

Synonymes: 2210418N07 Kit, Cas-1 Kit, Cas1 Kit, Cs-1 Kit, CS1 Kit, Cat01 Kit, Catl Kit, fb68a12 Kit, wu:fb68a12 Kit, CAT Kit, CATA Kit, CG6871 Kit, CT21282 Kit, CatA Kit, DMCATHPO Kit, DROCATHPO Kit, Dmel\\CG6871 Kit, U00145 Kit, bs36h11.y1 Kit, cat Kit, GB11648 Kit, catalase Kit, Cat Kit, BA0843 Kit, DKFZp469E0232 Kit, catalase Kit, Catalase Kit, catalase, gene 2 Kit, CAT Kit, Cat Kit, cat Kit, cat.2 Kit, LOC769804 Kit, BA_0843 Kit

Sujet: Quantitative determination of catalase activity by colorimetric (570nm) or fluorometric (530nm/590nm) methods.
Procedure: 40 min.

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. This improved assay directly measures catalase degradation of H2O2 using a redox dye. The change in color intensity at 570nm or fluorescence intensity (gamma em/ex = 585/530nm) is directly proportional to the catalase activity in the sample.