Catalase (CAT) Colorimetric Assay Kit

    • Biochemical Assay Kit-Elabscience
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    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K031-S

      Size:
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $120

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry (Visible range)

      Valid period: 6 months

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      General information

      Detection significance

      Catalase (CAT) is an enzyme in organism that can efficiently and specifically decompose hydrogen peroxide and is a binding enzyme with iron porphyrin as an auxiliary group. CAT clears hydrogen peroxide in the body and protects cells from the toxicity of H2O2. CAT can also oxidize certain cytotoxic substances, such as formaldehyde, formic acid, phenol and ethanol. According to the difference of catalytic center structure, CAT can be divided into two types, one is iron porphyrin structure, also known as iron porphyrin enzyme, the other contain manganese ion, also known as manganese catalase. CAT is common in breathing organisms. It is mainly found in chloroplasts, mitochondria, endoplasmic reticulum, liver and red blood cells of animals.

      Detection principle

      The reaction that catalase (CAT) decomposes H2O2 can be quickly stopped by ammonium molybdate. The residual H2O2 reacts with ammonium molybdate to generate a yellowish complex. CAT activity can be calculated by production of the yellowish complex at 405 nm.

      Operation procedures

      Operation steps

      1)    Control tube: Add 1 mL of Reagent 1 into the 5 mL EP tubes.

      Sample tube: Add a* mL of sample and 1 mL of Reagent 1 into the 5 mL EP tubes.

      2)    Incubate at 37℃ for 5 min.

      3)    Add 0.1 mL of Reagent 2 into each tube, mix fully and react at 37℃ for 1 min accurately.

      4)    Sample tube: Add 1 mL of Reagent 3 application solution and 0.1 mL of Reagent 4, mix fully.

      Control tube: Add 1 mL of Reagent 3 application solution, 0.1 mL of Reagent 4 and a* mL of sample, mix fully.

      5)    Stand for 10 min at room temperature. Set to zero with double-distilled water and measure the OD values of each tube at 405 nm with 0.5 cm diameter cuvette.

      Operation table

       

      Control tube

      Sample tube

      Sample (mL)

       

      a*

      Reagent 1 (mL)

      1.0

      1.0

      Mix fully and incubate at 37 for 5 min.

      Reagent 2 (mL)

      0.1

      0.1

      Mix fully and react accurately at 37 for 1 min.

      Reagent 3 (mL)

      1.0

      1.0

      Reagent 4 (mL)

      0.1

      0.1

      Sample (mL)*

      a*

       

      Mix thoroughly, stand for 10 min at room temperature. Set to zero with double-distilled water and measure the OD values of each tube at 405 nm with 0.5 cm diameter cuvette.

      Note: 1) If there are no obvious hemolysis and blood fat for serum (plasma) samples, or there is no high- fat in tissue homogenate samples, * can be replaced by double-distilled water for control tube. Control tubes only need 1-2 for each experiment.

      2) For Serum or plasma samples, a* is 0.1 mL. For Tissue homogenates, a* is 0.05 mL.

      Performance characteristics

      Technical parameter

      Detection range 0.27-155.4 U/mL Average inter-assay CV 5.1%
      Sensitivity 0.27 U/mL Average intra-assay CV 3.1%
      Average recovery rate 96%
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