Overview

  • Product name
    Total Antioxidant Capacity Assay Kit
  • Detection method
    Colorimetric
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Quantitative
  • Assay time
    2h 00m
  • Product overview

    Total Antioxidant Capacity Assay Kit ab65329 can measure either the combination of both small molecule antioxidants and proteins, or small molecules alone in the presence of our proprietary Protein Mask.


    In the total antioxidant capacity assay protocol, the Cu2+ ion is converted to Cu+ by both small molecule and protein antioxidants. The Protein Mask prevents Cu2+ reduction by proteins, enabling the analysis of only the small molecule antioxidants. The reduced Cu+ ion is chelated with a colorimetric probe giving a broad absorbance peak around 570 nm, proportional to the total antioxidant capacity.


    Total antioxidant capacity assay protocol summary:
    - add protein mask to samples if only measuring small molecule total antioxidant capacity
    - add samples and standards to wells
    - add Cu2+ solution and incubate for 90 min at room temp
    - analyze with microplate reader

  • Notes

    Antioxidants play an important role in preventing the formation of, and scavenging of, free radicals and other oxidizing species. There are three categories of antioxidant species: enzyme systems (GSH reductase, catalase, peroxidase, etc.), small molecules (ascorbate, uric acid, GSH, vitamin E, etc.) and proteins (albumin, transferrin, etc.). 

    Trolox is used to standardize antioxidants, with all other antioxidants being measured in Trolox equivalents. Measurement of the combined non-enzymatic antioxidant capacity of biological fluids and other samples provides an indication of the overall capability to counteract reactive oxygen species (ROS), resist oxidative damage and combat oxidative stress-related diseases.

    Related products

    Review the oxidative stress marker and assay guide to learn about more assays for oxidative stress.

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components Identifier 100 tests
    Assay Diluent WM 1 x 10ml
    Cu++ Reagent Blue 1 x 0.2ml
    Protein Mask NM 1 x 10ml
    Trolox Standard (1 µmole) (Lyophilized) Yellow 1 vial
  • Research areas
  • Relevance
    Antioxidants play an important role in preventing the formation of and scavenging of free radicals and other potentially toxic oxidizing species. There are three categories of antioxidant species: enzyme systems (GSH reductase, catalase, peroxidase, etc.), small molecules (ascorbate, uric acid, GSH, vitamin E, etc.) and proteins (albumin, transferrin, etc.). Different antioxidants vary in their reducing power, and cooperation of all different antioxidants provides greater protection against reactive oxigen or nitrogen radicals than any single compound alone. Therefore, the overall total antioxidant capacity may give more relevant biological information compared to that obtained by the measurement of individual components, as it considers the cumulative effect of all antioxidants present.

Images

  • Total antioxidant capacity was determined in mouse cardiac homogenates using Total antioxidant capacity assay kit (ab65329). 

  • Trolox equivalent capacity measured in milk and concentrated squash. Background signal subtracted, duplicates; +/- SD

  • Trolox equivalent capacity measured in mouse tissue lysates, showing quantity (nmol) per mg of extracted protein. Results following blocking of protein activity is shown (Mask). (Duplicates; +/- SD).

  • Trolox equivalent capacity measured in biological fluids. Results following blocking of protein activity is shown (Mask). Background signal subtracted, duplicates; +/- SD.

Protocols

References

This product has been referenced in:
  • Sacks J  et al. Effect of Roux-en-Y gastric bypass on liver mitochondrial dynamics in a rat model of obesity. Physiol Rep 6:N/A (2018). Functional Studies . Read more (PubMed: 29464885) »
  • Reddy KE  et al. Effects of High Levels of Deoxynivalenol and Zearalenone on Growth Performance, and Hematological and Immunological Parameters in Pigs. Toxins (Basel) 10:N/A (2018). Read more (PubMed: 29518941) »
See all 19 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Abreviews
Plant materials
Rice plants from control and Fe stress (300 ppm Fe2+ for 3 weeks) treatments were harvested with liquid nitrogen and stored in -80 °C freezer. Rice shoot tissues were applied in the test.
Experiment procedure
Grind the rice shoot tissues to fine powder with pre-chilled pestle and mortar in liquid N. Around 50 mg of samples were weighted in 2 ml micro-centrifuge tubes and 1 ml of PBS were added to extract the total antioxidants. After 2 mins of vigorously vortex, the mixtures were placed on ice for 10 mins. Subsequently, the tubes were centrifuged at 14,000 g for 5 mins at 4 °C followed by collecting the supernatants into fresh 1.5 ml micro-centrifuge tubes. To remove the small debris, the tubes were centrifuged again at the same speed for 2 mins.
Load the trolox standards and samples to the micro-plate according to the protocol. Both standards and samples were measured with two replicates. After adding the Cu2+ reagents, the micro-plate was incubated at room temperature in dark for 90 mins. Measure the absorbance of different wells at the wavelength of 570 nm.
Results
The total antioxidant capacity (trolox equivalent) was significantly induced by the excess Fe treatment in rice shoot tissue.

additional notes: ABTRIAL-TVX7B

Abcam user community

Verified customer

Submitted Jan 24 2018

Abreviews
Plant materials
Rice plants from control and Fe stress (300 ppm Fe2+ for 3 weeks) treatments were harvested with liquid nitrogen and stored in -80 °C freezer. Rice shoot tissues were applied in the test.
Experiment procedure
Grind the rice shoot tissues to fine powder with pre-chilled pestle and mortar in liquid N. Around 50 mg of samples were weighted in 2 ml micro-centrifuge tubes and 1 ml of PBS were added to extract the total antioxidants. After 2 mins of vigorously vortex, the mixtures were placed on ice for 10 mins. Subsequently, the tubes were centrifuged at 14,000 g for 5 mins at 4 °C followed by collecting the supernatants into fresh 1.5 ml micro-centrifuge tubes. To remove the small debris, the tubes were centrifuged again at the same speed for 2 mins.
Load the trolox standards and samples to the micro-plate according to the protocol. Both standards and samples were measured with two replicates. After adding the Cu2+ reagents, the micro-plate was incubated at room temperature in dark for 90 mins. Measure the absorbance of different wells at the wavelength of 570 nm.
Results
The total antioxidant capacity (trolox equivalent) was significantly induced by the excess Fe treatment in rice shoot tissue (see attached image).

Abcam user community

Verified customer

Submitted Oct 12 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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