Overview

  • Product name

    Ascorbic Acid Assay Kit (Fluorometric)
    See all Ascorbic acid kits
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate
  • Assay type

    Quantitative
  • Sensitivity

    1 µM
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Ascorbic Acid Assay Kit (Fluorometric) (ab219928) provides a sensitive fluorescent assay for quantifying total ascorbic acid in cell lysate. This assay is based on the enzymatic oxidation of ascorbic acid to dehydroascorbate acid (DHA). DHA can then in turn be detected by our Blue Dye with a fluorescence microplate reader at Ex/Em = 340/430 nm. The assay can detect 1µM of total ascorbate.


    This product can also be used to quantify the ratio of DHA to ascorbic acid. Under physiological conditions, DHA concentrations are extremely low. However, extracellular ascorbic acid is massively oxidized to DHA in certain neuropathological diseases, leading to an increase in the uptake of DHA in neurons, which can in turn result in neuronal damage.

  • Notes

    L-Ascorbic Acid (also called Vitamin C) is a critical metabolite for both plant and animals in cell division, growth and defense. Ascorbate is produced from glucose in the liver of most mammalian species. For humans ascorbate has to be obtained from food to survive, and a lack of sufficient Vitamin C can result in scurvy, and may eventually lead to death. As an antioxidant ascorbate can reduce the risk of developing chronic disease such as cancer and cardiovascular disease. In food industry, ascorbic acid and its sodium, potassium, and calcium salts are commonly used as antioxidant food additives to prevent undesired color and taste.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 200 tests
    Ascorbic Acid Standard (3.52 mg) 1 x 3.52mg
    Assay Buffer 1 x 20ml
    Blue Dye 1 vial
    DMSO 1 x 100µl
    Enzyme Mix 2 vials
  • 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.

Images

  • Typical Ascorbic Acid standard calibration curve. Ascorbic Acid dose response was measured on a solid black 96 well plate using a Gemini microplate reader (Molecular Devices). As low as 1 µM ascorbic acid can be detected with 30min incubation.

Protocols

References

ab219928 has not yet been referenced specifically in any publications.

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