Total Cholesterol (TC) Colorimetric Assay Kit (Single Reagent, COD-PAP Method)

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

      Catalog number:E-BC-K109-S

      Size:
      • 100Assays
      • 50Assays
      Qty:
      - +
      Price: $180

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometer

      Valid period: 6 months

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

      Add to cart Compare Bulk request Manual

      General information

      Detection significance

      Cholesterol is a kind of sterol and lipid in cell membrane. Most of cholesterol in blood exists in the form of cholesterol ester. Human plasma lecithin-cholesterol acyltransferase is an enzyme that catalyzes the formation of cholesterol ester. Cholesterol synthesized or deposited in peripheral cells returns to the liver through the reverse cholesterol transport system for reuse or re-conversion into bile acids.

      Detection principle

      Total cholesterol includes free cholesterol and cholesterol esters. Cholesterol ester can be hydrolyzed by cholesterol esterase to produce cholesterol and free fatty acid. Cholesterol is oxidized by cholesterol oxidase to produce △4-cholestenone and hydrogen peroxide. In the presence of 4-aminoamylpyridine and phenol, hydrogen peroxide catalyze peroxidase to form red quinone compounds of benzoquinone imine phenizone.

      The key point

      1.       Protect the reagent from contamination of glucose, cholesterol, etc.

      2.       The samples should not contain reducing substances such as ascorbic acid or glutathione, otherwise the generated hydrogen peroxide will be consumed, and competitive      inhibition will result in low measurement results.

      3.       The amount of sample and reagent can be increased and decreased as the ratio of 1:100.

      Operation procedures

      Operation steps

      1.    Blank tube: Add 10 μL of Double distilled water into a 2 mL EP tube.

      Standard tube: Add 10 μL of reagent 2 into a 2 mL EP tube.

      Sample tube: Add 10 μL of Sample into a 2 mL EP tube.

      2.    Add 1000 μL of Reagent 1 and mix fully.

      3.    Incubate at 37 for 10 min. Set to zero with double distilled water and measure the OD values of each tube at 550 nm with 0.5 cm diameter cuvette.

      Operation table


       

      Blank tube

      Standard tube

      Sample tube

      Double distilled water (μL)

      10

       

       

      Standard (μL)

       

      10

       

      Sample (μL)

       

       

      10

      Working solution (μL)

      1000

      1000

      1000

      Mix thoroughly, incubate at 37 for 10 min. Set spectrophotometer to zero with distilled water and measure the OD value at 510 nm with 0.5 cm diameter cuvette.

      Performance characteristics

      Technical parameter

      Detection range 0.09-25.85 mmol/L Average inter-assay CV 2.8%
      Sensitivity 0.09 mmol/L Average intra-assay CV 1.1%
      Average recovery rate 102%

      Citations

      1. Alagebrium Mitigates Metabolic Insults in High Carbohydrate High Fat Diet Fed Rats
        Journal: Pharmaceutical Sciences(2019)
        DOI: 99999999
        Products: E-BC-K109-S E-BC-K261-S 
        Sample type: Serum
        Reactivity: Rat
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