Abcam, ab65656, Ascorbic Acid Assay Kit (Biological Samples)

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Ascorbic Acid Assay Kit (Colorimetric) (ab65656) uses an iron reduction method in combination with ascorbate oxidase to quantify ascorbic acid (vitamin C, ascorbate) levels. Readout on any colorimetric (545-600nm) plate reader. Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Cell culture extracts, Tissue Extracts, Cell culture media, Cell culture supernatant, Serum, Other biological fluids,
Assay type:Quantitative,
Sensitivity:> 0.2 nmol/well,
Range:0.2 - 20 nmol/well,
Assay time:30m,
Assay Platform:Microplate reader

Product details:
Ascorbic Acid Assay Kit (Colorimetric) (ab65656) provides a rapid, simple, and sensitive means of detecting ascorbic acid (vitamin C) in biological samples such as serum and other body fluids, tissue and cell extracts, growth media and food products.
Ascorbic assay principle
In the ascorbic acid assay protocol, 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.
Ascorbic acid assay protocol summary
- add samples and standards to wells
- add reaction mix
- analyze with a microplate reader in kinetic mode every minute for 2-3 minutes
Related Ascorbic Acid assays
- We recommend using this kit ab65656 particularly for plasma and serum samples
- For other sample types, we recommend Ascorbic Acid Assay Kit (Colorimetric/Fluorometric)
ab65346
, which is based on a non-enzyme catalyst method and is slightly simpler to use than this kit as it does not use the additional ascorbate oxidase treatment step
- Ascorbic Acid Assay Kit (Fluorometric)
ab219928
uses an alternative method based on ascorbate oxidase.
Other Notes
This product was previously called K671 Biovision Ascorbic Acid Colorimetric Assay Kit II (FRASC). Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Ascorbic acid also known as vitamin C plays an important mechanical role as an antioxidant and a cofactor in various enzymatic reactions. It is water-soluble and has a molecular mass of approximately 176.12 g/mol. Ascorbic acid is widely expressed in fruits and vegetables and cells in both animals and plants. Inside cells it primarily exists as ascorbate which can undergo oxidation by the enzyme ascorbate oxidase relevant in some assay tools used in biology research.
Biological function summary
As a potent reducing agent ascorbic acid is critical for the hydroxylation of collagen which is essential for connective tissue strength in humans and animals. It also participates in the synthesis of carnitine involved in fat metabolism and acts in the biosynthesis of neurotransmitters. Ascorbic acid forms a part of antioxidant defense complexes working alongside enzymes like glutathione peroxidase to protect cells against oxidative stress.
Pathways
Ascorbic acid is integral in the citrate cycle (TCA cycle) and the pentose phosphate pathway. It interacts with proteins such as prolyl hydroxylase involved in collagen synthesis within the TCA cycle. The pentose phosphate pathway helps produce NADPH which works with ascorbic acid in maintaining cellular antioxidant status supporting its role in the reduction of oxidized forms of essential biomolecules.
Ascorbic acid deficiency leads to scurvy a condition characterized by weakened connective tissue leading to symptoms like bleeding gums and joint pain. It also plays a role in diabetes with oxidative stress being an underlying mechanism linked to the disease. Ascorbic acid's connection to oxidative damage involves proteins like advanced glycation end-products found in diabetic complications where its antioxidant properties help mitigate related tissue damage.