Ascorbic Acid Colorimetric/Fluorometric Assay Kit

Catalog #: K661 | abID: ab65346

Product Details

abID ab65346
Cat # +Size K661-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The assay can detect 0.01-10 nmol of ascorbic acid per assay in various samples.
Features & Benefits • Simple procedure; takes ~ 5 minutes for the reaction to complete
• Fast and convenient
Kit Components • Ascorbic Acid Assay Buffer
• Ascorbic Acid Probe (in DMSO)
• Catalyst
• Ascorbic Acid Enzyme Mix (lyophilized)
• Ascorbic Acid Standard (20 µmole)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


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 is a wide variety of foods and biological specimens. It is important to be able to monitor ascorbic acid content in these different samples. BioVision’s Ascorbic Acid Assay Kit provides a rapid, simple, and sensitive means of detecting ascorbic acid in various biological samples. In this assay, our proprietary catalyst oxidizes ascorbic acid to produce a product that interacts with the ascorbic acid probe, generating color and fluorescence. Ascorbic acid can be easily determined by either colorimetric (spectrophotometry at λ = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods. The assay can detect 0.01-10 nmol of ascorbic acid per assay in various samples.

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Can carrots or berries be used as samples for this kit?
Simply homogenize the carrots/berries using the assay buffer in the kit and then centrifuge at top speed in a microcentrifuge to pellet debris. Collect the supernatant for the assay. It will be important to optimize the sample volume such that the solution in the well does not become red/pink since OD 570 nm is in the pink-red range of the spectrum.
Does the kit quantify only the intact vitamin C or can it distinguish between intact and oxidized vitamin C?
In this assay, our proprietary catalyst oxidizes ascorbic acid to produce a product that interacts with the ascorbic acid probe, generating color and fluorescence. So if the vitamin C is already oxidized, it is likely to generate color/fluorescence in the reaction without the catalyst. This value can then be subtracted from the +catalyst reaction readings for the same sample so that the amount of un-oxidized vitamin C can be calculated.
Can this kit be used for serum ascorbic acid estimation?
Due to interference from other proteins and serum components we recommend using the K671 for Serum Ascorbic acid quantification.
Would your kits K661-100 or K671-100 be suitable for measuring ascorbic acid levels in blackcurrant juice? The juice itself has quite a strong colour. Even with diluting it down, would you know if one of these 2 kits could be suitable?
K661 and K671 measurements are in the yellow-orange range of the spectrum (545-600nm). There could be some interference from the blackcurrant juice in this range but you can run parallel controls and subtract the background to get the final numbers for your assay. So either one of these could work. K661 allows both colorimetric and fluorometric measurements and K671 allows only colorimetric measurement. I would recommend not using the fluorometric version of the assay since it is 10-100 fold more sensitive and the background color intereference may be more pronounced in this case.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples since this is an enzyme activity assay. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Cui, J el. Al. Progressive Pseudogenization: Vitamin C Synthesis and Its Loss in Bats. Mol. Biol. Evol., Feb 2011; 28: 1025 - 1031.
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