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    total-antioxidant-capacity-assay-kit-ab65329.pdf

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Total Antioxidant Capacity Assay Kit (ab65329)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (3)Q&A (34)References (54)

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Functional studies - ab65329
  • Functional assays: TAC (ab65329)
  • Functional assays: TAC (ab65329)
  • Functional assays: TAC (ab65329)
  • Typical Trolox standard calibration curve

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 2 hr
  • Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine

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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

    This product is manufactured by BioVision, an Abcam company and was previously called K274 Total Antioxidant Capacity (TAC) Colorimetric Assay Kit. K274-100 is the same size as the 100 test size of ab65329.

    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 µmol) (Lyophilized) Yellow 1 vial
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Oxidative stress
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Oxidative Stress Assay Kits
    • Oxidative Stress
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Antioxidant
  • 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

  • Functional studies - ab65329
    Functional studies - ab65329Image from Wan X et al., PLoS Pathog 12(10), Fig 5D. doi: 10.1371/journal.ppat.1005954 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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

  • Functional assays: TAC (ab65329)
    Functional assays: TAC (ab65329)

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

  • Functional assays: TAC (ab65329)
    Functional assays: TAC (ab65329)

    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).

  • Functional assays: TAC (ab65329)
    Functional assays: TAC (ab65329)

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

  • Typical Trolox standard calibration curve
    Typical Trolox standard calibration curve

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (54)

Publishing research using ab65329? Please let us know so that we can cite the reference in this datasheet.

ab65329 has been referenced in 54 publications.

  • Madariaga VI  et al. Myogenous temporomandibular disorders and salivary markers of oxidative stress-A cross-sectional study. J Oral Rehabil 48:1-9 (2021). PubMed: 32979853
  • Li M  et al. Brachyury engineers cardiac repair competent stem cells. Stem Cells Transl Med 10:385-397 (2021). PubMed: 33098750
  • Okda TM  et al. Chemopreventive and anticancer activities of indomethacin and vitamin D combination on colorectal cancer induced by 1,2-dimethylhydrazine in rats. Biomed Rep 14:27 (2021). PubMed: 33408861
  • Shili CN  et al. Effect of a Phytogenic Water Additive on Growth Performance, Blood Metabolites and Gene Expression of Amino Acid Transporters in Nursery Pigs Fed with Low-Protein/High-Carbohydrate Diets. Animals (Basel) 11:N/A (2021). PubMed: 33672517
  • D'Onofrio N  et al. Phenolic Profiles of Red Wine Relate to Vascular Endothelial Benefits Mediated by SIRT1 and SIRT6. Int J Mol Sci 22:N/A (2021). PubMed: 34073604
View all Publications for this product

Customer reviews and Q&As

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31-37 of 37 Abreviews or Q&A

Question

hi. I did my experiments using this kit. I have few queries regarding the calculation: 1. If I represents my results in terms of 'Antioxidant Capacity', what will be the units. 2. or if I use Sa/Sv formula (the second way to calculate the results give in the product insert). As Sv is the undiluted sample volume added per well, suppose if I use two different samples with different protein concentration (one is 5ug/ul and other is 30ug/ul) and add 10 ul in one well and other 50 ul in other well and then make up final volume to 100. The results will automatically be very much different. Is it correct to calculate the same, one may get biased to the samples. So, in that case what i did, i used same same conc of protein samples (say 25ug for each sample) irrespective of undiluted sample volume and made to the final 100ul volume. Is it ok to do that way? Just want your suggestion. As I think, if one will calculate the results by volume taken form undiluted aliquot, the results will be bised. waiting for your quick response. Bes

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Abcam community

Verified customer

Asked on Oct 22 2011

Answer

Thank you for contacting Abcam about ab65329. The answers to your questions are below:        1.    The units of measurement will be in nmols/ul or in mM of Trolox (i.e. the equivalent that                 you will the comparing the sample to). Taking the same amount of protein in each sample will work as it will be you absolute comparative results. In that case as well you will have to keep a note on the sample volume that is being used (will be different for each sample) in order to calculate the TAC. Please let me know if there is anything else I can help you with.  

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Abcam Scientific Support

Answered on Oct 22 2011

Question

Hi, I want to order Total Antioxidant Capacity Assay Kit (ab65329), but before that I have few queries. I want to measure the TAC of my protein. Can I perform this assay for a GST tagged protein or not?. Will there be any interference with DTT or not? Hoping for a quick response at your end. best

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Abcam community

Verified customer

Asked on Oct 05 2011

Answer

Thank you for your original request for information and your patience while I communicated with the laboratory. They informed that we have not tested the interference of this kit with DTT; however it should be very compatible with the indicated sample type. Due to the fact that the affect of DTT has not been determined we cannot guarantee that this kit will work for your experiment. Please let me know if you have any more questions.

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Abcam Scientific Support

Answered on Oct 05 2011

Question

Is ab65329 able to measure antioxidant enzyme as well as antioxidant proteins and small molecules?

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Abcam community

Verified customer

Asked on Jun 14 2011

Answer

Thank you for your enquiry. With this kit one can measure either the combination of both small molecule antioxidants and proteins or small molecules alone. This kit does not measure antioxidant enzyme systems. I hope this is helpful. Please contact us again if you need any further information.

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Abcam Scientific Support

Answered on Jun 14 2011

Question

Is this kit suitable for use with rat samples?

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Abcam community

Verified customer

Asked on Apr 11 2011

Answer

Thank you for contacting us. Yes, this kit should be compatible with rat samples. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Apr 11 2011

Question

I need a few information about price and the others information for your kit, Total Antioxidant Capacity. Can I be using this Kit for detection of antioxidant in human serum? I have been collecting human blood - EDTA as anticoagulant, and I want to check the antioxidant capacity in the samples. If I cannot use the Kit for my samples (serum-EDTA), do you have the other kit for detection antioxidant status? Please, give me information the procedure and the price. Thank you for your kindness.

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Abcam community

Verified customer

Asked on Oct 15 2010

Answer

Thank you for your enquiry. Please rest assured that this product can be used for human serum samples, urine, culture media, food and drinks. Also, no sample purification from these sources is necessary. However, please bear in mind that the EDTA anticoagulant will give a lower antioxidant capacity compared with heparinized plasma samples but nonetheless, sufficient for detection and data analysis. I have included a publication mentioning the difference between EDTA and heparin anticoagulants for your reference. The effect of anticoagulant choice on apparent total antioxidant capacity using three different methods. Ann Clin Biochem. 1995 Jul;32 ( Pt 4):413-6. http://www.ncbi.nlm.nih.gov/pubmed/7486802 The selling price for this product is 85,000 yen. However, the price may be slightly lower if you purchase from your local sub-dealer. Please consult them for more information on price and purchasing. Lastly, the protocols for the kit are quite straight forward and can be referred to on the datasheet under [Instructions For Use/Protocol] link. I hope this information will be helpful.

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Abcam Scientific Support

Answered on Oct 15 2010

Question

Dear Sir or Madam, I have ordered a total antioxidant capacity kit (4x). I have 2 questions to this regard: - Does this kit work with both EDTA- and Citrate plasma? - What is the current shipping status? Thank you for your assistance. Sincerely,

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Abcam community

Verified customer

Asked on Jun 08 2010

Answer

Thank you for your enquiry and your interest in our products. I can confirm that this kit (Total Antioxidant Capacity Assay Kit) will work with plasma samples with trace amounts of EDTA/ Citrate. You should not have any issues. Currently, this kit is available and we have 3 in our UK stock. I hope this helps and if I can assist further, please do not hesitate to contact me.

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Abcam Scientific Support

Answered on Jun 08 2010

Question

May I kindly enquire for the below mentioned products, can they be tested for Saliva / Serum? ab65354 Superoxide Dismutase Activity Colorimetric Assay Kit 100 tests ab65329 Total Antioxidant Capacity Assay Kit 100 tests ab65344 Uric Acid Assay Kit 100 tests ab65656 Ascorbic Acid Assay Kit (Biological Samples) 100 tests I guessed for ab65656, it is mentioned that it can be used for serums and fluids. Please get back to me asap. Thanks alot!

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Abcam community

Verified customer

Asked on Jul 30 2008

Answer

Thank you for your enquiry. I can confirm that in the [Instructions for use/protocol] section of the datasheet, it states that these products (ab65329, ab65344, ab65656) can be used with serum samples. Although product ab65354 does not have the same information, it is recommended for use in detecting SOD activity in tissue homogenates, cell lysates, and other biological fluids/extracts where forms of SOD might be present. I hope this information will be helpful. If you should require any other information, please do not hesitate to contact me.

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Abcam Scientific Support

Answered on Jul 30 2008

31-37 of 37 Abreviews or Q&A

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