• Product name
    Ascorbic Acid Assay Kit (Biological Samples)
    See all Ascorbic acid kits
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media
  • Assay type
  • Sensitivity
    > 0.2 nmol/well
  • Range
    0.2 nmol/well - 20 nmol/well
  • Assay time
    0h 30m
  • Product overview

    Ascorbic Acid Assay Kit (Biological Samples) (ab65656) provides a rapid, simple, and sensitive means of detecting ascorbic acid in biological samples such as serum and other body fluids, tissue and cell extracts, growth media and food products. In this assay, Fe3+ is reduced to Fe2+ by any antioxidants present. The ferrous iron is chelated with a colorimetric probe to produce a product with a strong absorbance band which can be monitored between OD=545-600 nm. The addition of ascorbate oxidase to parallel samples removes any ascorbate present leaving a background value which is subtracted from the total to give ascorbate content. The assay can detect 0.2 to 20 nmol of ascorbic acid in various samples.
    Visit our FAQs page for tips and troubleshooting.


    Suitable for biological samples Range: 0.2 to 20 nmol of ascorbic acid

  • Notes

    Ascorbic Acid (Vitamin C) plays an important role in many biological processes. It is a potent anti-oxidant, anti-inflammatory, anti-viral agent and an immune stimulant and is present in a wide variety of biological specimens. Due to the presence of a variety of other antioxidants in biological samples such as serum, most ascorbic acid assays show strong interference.

  • Tested applications
    Suitable for: Functional Studiesmore details


  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Ascorbate Oxidase (lyophilized) 1 vial
    Ascorbic Acid Probe 1 x 1ml
    Ascorbic Acid Standard (Lyophilised) 1 vial
    FeCl3 solution 1 x 1ml
    Ferric Reducing Ascorbate Buffer 1 x 25ml
  • Research areas
  • Relevance
    Ascorbic Acid (C6H8O6) is an essential nutrient in human diets, and is necessary to maintain connective tissue and bone. The biologically active form of ascorbic acid is vitamic C, a water soluble vitamin which is the L-enantiomer form of ascorbic acid. Humans have lost the ability to synthesize ascorbic acid and must obtain in the diet. Ascorbic acid is an electron donor for enzymes involved in collagen hydroxylation, biosynthesis of carnitine and norepinephrine, tyrosine metabolism and amidatin of peptide hormones. The ability of vitamin C to donate electrons also makes it a potent water-soluble anti-oxidant that readily scavenges free radicals.


Our Abpromise guarantee covers the use of ab65656 in the following tested applications.

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

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.


  • Standard curve : mean of duplicates (+/-SD) with background readings subtracted



This product has been referenced in:
  • Blaschke K  et al. Vitamin C induces Tet-dependent DNA demethylation and a blastocyst-like state in ES cells. Nature 500:222-6 (2013). Read more (PubMed: 23812591) »
  • Dai D  et al. Healing of saccular aneurysms following platinum coil embolization: lack of improved efficacy with vitamin C supplementation. J Neurointerv Surg N/A:N/A (2012). Functional Studies ; Rabbit . Read more (PubMed: 22914744) »

See all 2 Publications for this product

Customer reviews and Q&As

Unfortunately, this kit will measure the level of Vitamin C, Ascorbic Acid, in biological samples, but not the protein you are after. Ascorbate peroxidases (or APX1) are enzymes that detoxify peroxides such as hydrogen peroxide using ascorbate as ...

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I am sorry for the delay in getting back to you. I now have further input from the lab.
From the results you have shared it does seem like there is higher background in your samples than would be expected. This can often be improved by optimizing t...

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Thank you for contacting us yesterday in regards to the Ascorbic Acid Assay Kit (ab65656). I was sorry to hear you have been having some difficulties in achieving consistent results with your samples.
As discussed over the phone, I would like to t...

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Thank you for contacting us.

The lab confirmed thata 0.1% SDS based lysis buffer can be used for cell lysis, and would be compatible with the kit.

I hope this information is helpful to you. Please do not hesitate to contact us if ...

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Thank you for contacting us. Because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products and kits.

We are happy to reassure our customers that all of our products are covered by our Abpromise, w...

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Thank you for your email. Each kit contains sufficient reagents for 100 tests, therefore it is possible to test ~40 samples in duplicate with each kit (6 data points in the standard curve). Therefore you will require 3 kits for 120 samples. ...

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Thank you for your patience. Yes, I can confirm that in the kit, FeCl3 is present in sufficient amount that EDTA (used at the concentration as anti-coagulant in plasma/serum samples) will not interfere with the assay. One would need to use a lot ...

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Thank you for your inquiry. I just heard back from the testing laboratory that these kits cannot distinguish ascorbic acid isoforms. I am sorry that this information could not be more helpful, but please let us know if we can be of any other help.

Thank you for contacting us. I have hear back from the ab with the following information to your questions: 1. Yes, you can use plasma samples – but for ab65656 we recommend removing the proteins (e.g. by using spin filter (10 kDa cutoff) (ab933...

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Thank you for contacting us. We do not have data from tissue samples other than serum but we assume the kit will be suitable for these samples. You will need to determine how much sample to add to keep the optical densities within the linear range ...

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