Total Antioxidant Capacity (T-AOC) Colorimetric Assay Kit

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

      Catalog number:E-BC-K136-S

      Size:
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $150

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry (Visible range)

      Valid period: 6 months

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

      Add to cart Compare Bulk request Manual

      Application

      The kit is used for the determination of total antioxidant capacity (T-AOC) in serum, plasma, tissue, cell, whole blood samples.

       

      Detection principle

      A variety of antioxidant macromolecules, antioxidant molecules and enzymes in a system can eliminate all kinds of reactive oxygen species and prevent oxidative stress induced by reactive oxygen species. The total level reflect the total antioxidant capacity in the system. Many antioxidants in the body can reduce Fe3+ to Fe2+ and Fe2+ can form stable complexes with phenanthroline substance. The antioxidant capacity (T-AOC) can be calculated by measuring the absorbance at 520 nm.

       

      Experimental instrument

      Micropipettor, Vortex mixer, Thermostat incubator, Centrifuge, Spectrophotometry (520 nm), Balance.

       

      Sample pretreatment

      1.    Serum (plasma) samples:

      For blood samples, timely separate serum or plasma from blood after collection and avoid hemolysis. For cell supernatant and other liquid samples, directly collect samples for measurement. It is recommended to use heparin or sodium citrate as anticoagulation, not EDTA.

       

      2.    Tissue samples

      Weigh the tissue accurately. Add 9 times the volume of normal saline or PBS according to the ratio of Weight (g): Volume (mL) =1:9. Mechanical homogenate the sample in ice water bath to prepare 10% homogenate. Centrifuge at 4ºC at 3500 rpm for 5 min and collect the supernatant for measurement. Meanwhile, determine the protein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165).

       

      3.    Whole blood samples

      Add 9 times the volume of ddH2O according to the ratio of Weight (g): Volume (mL) =1:9 to prepare 10% hematolysis.

       

      4.    Cell samples

      Collect 1×106 cells with cell scraper (Don’t use trypsin or EDTA). Add 400 μL of cold PBS(0.01M, pH7.4), then treat the sample with mechanical homogenate or sonication on ice. Centrifuge at 4ºC at 4000 rpm for 15 min and collect the supernatant for measurement. Meanwhile, determine the protein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165).

       

      Operation steps

      1.    Serum (plasma) samples


       

      Sample tube

      Control tube

      Reagent 1 (mL)

      1.0

      1.0

      Sample (mL)

      0.1

       

      Reagent 2 working solution (mL)

      2.0

      2.0

      Reagent 3 working solution (mL)

      0.5

      0.5

      Mix fully, react at 37 for 30 min.

      Reagent 5 (mL)

      0.1

      0.1

      Sample (mL)

       

      0.1

      Mix fully and stand for 10 min. Set to zero with ddH2O and measure the OD values of each well at 520 nm with 1 cm cuvette.

       

      2.     Tissue samples


       

      Sample tube

      Control tube

      Reagent 1 (mL)

      1.0

      1.0

      10% Tissue homogenate (mL)

      a*

       

      Reagent 2 working solution (mL)

      2.0

      2.0

      Reagent 3 working solution (mL)

      0.5

      0.5

      Mix fully, react at 37 for 30 min.

      Reagent 5 (mL)

      0.2

      0.2

      10% Tissue homogenate (mL)

       

      a*

      Reagent 6 (mL)

      0.2

      0.2

      Mix fully and stand for 10 min. Set to zero with ddH2O and measure the OD values of each well at 520 nm with 1 cm cuvette.

      Note: It is recommended that a* is 0.1-0.2 mL.

       

      3.    Whole blood and cell samples

       

      Sample tube

      Control tube

      Reagent 1 (mL)

      1.0

      1.0

      Samples (mL)

      a*

       

      Reagent 2 working solution (mL)

      2.0

      2.0

      Reagent 3 working solution (mL)

      0.5

      0.5

      Mix fully, react at 37 for 30 min.

      Reagent 5 (mL)

      0.2

      0.2

      Samples (mL)

       

      a*

      Mix fully and stand for 10 min. Set to zero with ddH2O and measure the OD values of each well at 520 nm with 1 cm cuvette.

      Note: It is recommended that a* is 0.05 mL for whole blood and 0.1-0.2 mL for cell samples.

       

      Technical parameter

      1.    The sensitivity of the kit is 0.16 U/mL.

      2.    The intra-assay CV is 2.69% and the inter-assay CV is 8.23%.

      3.    The recovery of the kit is 105.13%.

      4.    The detection range of the kit is 0.16-145.2 U/mL.

       

      Notes

      1.  The kit is for scientific research only.

      2.  Instructions should be followed strictly, changes of operation may result in unreliable results.

      3.  The validity of kit is 6 months.

      4.  Do not use components from different batches of kit.


      • Show all (3)
      • Reviews (0)
      • Q&A (3)
      Q

      S****tSubmitted [ Mar 08 2019 ]

      Asked: This kit can use invertebrate hemolymph?

      Reply

      A

      adminSubmitted [ Mar 08 2019 ]

      Answered: Dear Serhat, Yes, the kit can use invertebrate hemolymph, if there is other question, please feel free to ask me. Regards, Cathy

      Q

      fSubmitted [ Dec 01 2018 ]

      Asked:  hi is this elisa kit suitable for detection of total antioxidant capacity in small intestine tissue of rat thank you

      Reply

      A

      adminSubmitted [ Dec 01 2018 ]

      Answered: This kit can dectect small intestine tissue sample.

      Q

      F***hSubmitted [ Nov 30 2018 ]

      Asked: Hi I want to ask if I want to measure Total antioxidant capacity in small intestine tissue of rat and number of test I require 75 sample can I use this kit and how many kit I require if I use kit for 100 assay thank you

      Reply

      A

      adminSubmitted [ Nov 30 2018 ]

      Answered: This kit of 100 assays could detect 50 samples with no duplication. So you need 2 kits of 100 assays if you won't test the samples in duplication.

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