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Oxaloacetate Colorimetric/Fluorometric Assay Kit

Catalog #: K659
$395.00

Product Details

Cat # +Size K659-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The Oxaloacetate Assay Kit can detect 0.1-10nmol (2-200 µM) of OAA.
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of OAA levels
Kit Components • OAA Assay Buffer
• OAA Probe ( in DMSO)
• OAA Enzyme Mix
• Developer
• OAA Standard (10 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Oxaloacetate (OAA, HOOC-CO-CH₂-COOH) is a TCA cycle intermediate. It precedes citrate which is formed by the transfer of an acetyl group to OAA. OAA is formed by the deamidation of aspartate or condensation of CO2 with pyruvate or PEP. Since mammals do not possess the enzymatic machinery to form TCA cycle intermediates from acetyl CoA, OAA is one of the anaplerotic entry points via pyruvate and pyruvate carboxykinase. BioVision's Oxaloacetate Assay Kit provides a simple, sensitive and rapid means of quantifying OAA in a variety of samples. In the assay, OAA is converted to pyruvate which is utilized to convert a nearly colorless probe to an intensely colored (λmax= 570nm) and fluorescent (Ex/Em = 535/587nm) product. The Oxaloacetate Assay Kit can detect 0.1-10nmol (2-200 µM) of OAA.


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Why is deproteinizing necessary for this assay?
We suggest deproteinizing samples using a PCA/KOH protocol (BioVision, Cat. #K808-200) or passing the samples through 10 kDa molecular weight cut off spin columns (BioVision, Cat # 1997-25). This will get rid of much of the protein in the samples and can remove the enzymes which can be converting OAA to pyruvate already.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. OAA is conserved through evolution. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
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.
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.