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Fumarate Colorimetric Assay Kit

Catalog #: K633
$425.00

Product Details

Cat # +Size K633-100
Size 100 assays
Detection Method Absorbance (450 nm)
Species Reactivity Mammalian
Applications Kit can detect as low as 1 nmol of fumarate per well (20 µM).
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
Kit Components • Fumarate Assay Buffer
• Fumarate Enzyme Mix
• Fumarate Developer
• Fumarate Standard (0.1 M)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Fumarate (HO₂CCH=CHCO₂H-) is an intermediate in the Kreb’s cycle used by cells to metabolize food to form ATP. In the mammalian liver, Fumarate is also a product of the Urea cycle where its release in the cytosol leads to its conversion into malate and subsequently oxaloacetate while generating NADH in the cytosol. The human skin naturally produces fumaric acid when exposed to sunlight. In fact, fumaric acid esters have been used to treat psoriasis, possibly due to an impaired production of fumaric acid in the skin. Fumaric acid has also been used in beverages, baking powders and candy. BioVision’s Fumarate Assay Kit provides a convenient tool for sensitive detection of the fumarate in a variety of samples. The fumarate Enzyme Mix recognizes fumarate as a specific substrate leading to proportional color development. The amount of fumarate can therefore be easily quantified using a colorimetric assay (λ = 450 nm). It can detect as low as 1 nmol of fumarate per well (20 µM).


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Immediately after the Fumarate Developer reconstitution with 0.9 ml of water the solution was strongly orange. There are no differences In the standard readings.
The developer could have been oxidized due to exposure to air/moisture. Also, it is important to reconstitute the developer in molecular grade deionized water. If regular water is used this can affect the function of the developer and contaminants in water can react with the developer. Either of these could lead to all orange color in all wells.
Can this kit be used to measure fumarate in microbial cultures?
Although this kit can be theoretically used with microbial samples, we have not tested this and hence it might need some tweaking of the protocol at the user’s end to achieve the best results.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. 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.
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.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. It is recommended to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
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.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.