製品の概要

  • 製品名
    Total Antioxidant Capacity Assay Kit
  • サンプルの種類
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • アッセイタイプ
    Quantitative
  • 全工程の試験時間
    2h 00m
  • 製品の概要

    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. Cu2+ ion is converted to Cu+ by both small molecule and protein. The Protein Mask prevents Cu2+ reduction by protein, 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.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    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. 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. In some cases, the antioxidant contribution of proteins is desired whereas in other cases only the contribution of the small molecule antioxidants is needed.

  • アプリケーション
    適用あり: Functional Studiesmore details

製品の特性

  • 関連性
    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.

アプリケーション

Our Abpromise guarantee covers the use of ab65329 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent dilution.

Total Antioxidant Capacity Assay Kit 画像

  • 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.

プロトコール

Total Antioxidant Capacity Assay Kit (ab65329) 使用論文

This product has been referenced in:
  • Truong T  et al. Comparative Ploidy Proteomics of Candida albicans Biofilms Unraveled the Role of the AHP1 Gene in the Biofilm Persistence Against Amphotericin B. Mol Cell Proteomics 15:3488-3500 (2016). Read more (PubMed: 27644984) »
  • Wan X  et al. SIRT1-PGC1a-NF?B Pathway of Oxidative and Inflammatory Stress during Trypanosoma cruzi Infection: Benefits of SIRT1-Targeted Therapy in Improving Heart Function in Chagas Disease. PLoS Pathog 12:e1005954 (2016). Human . Read more (PubMed: 27764247) »

See all 10 Publications for this product

Product Wall

We are currently investigating the problem as it is a very unusual complaint.

We have one last question; have you made Cu++ and other reagent fresh when you repeated the experiment or you just made them once and used again and again.

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Thank you for your enquiry.

Generally, serum can be stored at -20°C or at -70°. Freeze the tubes of serum rapidly, and store the aliquots protected from light. After thawing a tube, do not refreeze the serum unless you intend to store...

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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).
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Verified customer

投稿 Oct 12 2017

Plasma collected in glucose tubes will be compatible with the assay. There is no known interference from sodium fluoride or potassium oxalate.



The only real need for using Triton is if you are having difficulty homogenzing the cells in water, otherwise it is not necessary.



Yes, the kit can be used with samples from chicken tissues. We have not generated or received chicken data but the assay is suitable for any species. For an example of data from mouse heart tissue, please see figure 4 of the following reference:
...

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Unopened kit as received from Abcam can be stored for 6 months at 4C. Once reconstituted the standard should be stored at -20C. Please refer to the protocol booklet for further information.

If the total antioxidant capacity is desired, I would not recommend the use of the protein mask. If only the levels of the small molecule antioxidants are required, please use the protein mask.

Thank you for contacting us.

Although I don’t think EDTA will have an adverse effect on the efficiency of this kit, I would not recommend using EDTA for plasma preparations. Instead use heparin or citrate.

EDTA anticoagulant w...

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This kit has only been optimized for mammalian sample. Theoretically, it could be used in samples of multiple species including plant. Because we have not tested it with any plant sample, I am sorry that there is no specific protocol for measuring plan...

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

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