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Alpha-Ketoglutarate Colorimetric/Fluorometric Assay Kit

based on 6 citations in multiple journalsAlpha-Ketoglutarate Colorimetric/Fluorometric Assay Kit64.2 4
Highly Sensitive Assay
Catalog #: K677
$415.00

Product Details

Cat # +Size K677-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The α-Ketoglutarate Assay Kit is useful for detecting α-ketoglutarate in the range of 0.01 to 10 nmoles.
Features & Benefits • Simple procedure; takes ~ 30 minutes to complete the assay
• Fast and convenient
• Highly sensitive assays for measuring alpha-Ketaglutarate levels in biological samples
Kit Components • α-KG Assay Buffer
• α-KG OxiRed Probe (in DMSO)
• α-KG Converting Enzyme
• α-KG Development Enzyme Mix
• α-KG Standard (10 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

α-Ketoglutarate (α-KG) is a key intermediate in the Krebs cycle, coming after isocitrate and before succinyl CoA. Anaplerotic reactions replenish the cycle by synthesizing α-KG from transamination of glutamate, or through the action of glutamate dehydrogenase. α-KG is an important nitrogen transporter. Being a key intermediate, it is one of the organic acids measured in newborns as an indicator of inborn errors of metabolism. BioVision's α-Ketoglutarate Assay Kit provides a simple, sensitive and rapid means for quantifying α-KG in a variety of samples. In the assay, α-ketoglutarate is transaminated with the generation of pyruvate which is utilized to convert a nearly colorless probe to both color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The α-Ketoglutarate Assay Kit is useful for detecting α-ketoglutarate in the range of 0.01 to 10 nmoles.


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Can PCA precipitated supernatant be directly used in this assay?
You need to neutralize the sample before you use it for this assay.
Is the standard curve and the kit itself only accurate in the 0 - 10 nmol/sample range, or can the curve be extrapolated? That is, if the caculated value for a sample says 15 nmol, is that accurate, or do I need to dilute my samples more?
We recommend diluting the sample to fit within the std. curve linear range.
Will low concentrations of NADH in the sample interfere in this assay?
Theoretically, NADH should not interere. Pyruvate can generate background in the assay. The endogenous pyruvate interference can be subtracted from the sample reading using the parallel background control. Ideally the relative amount of pyruvate < amount of alpha KG in the sample.
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 times (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
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.
Liu, F. et al., (2017) MicroRNA-200c exacerbates the ischemia/reperfusion injury of heart through targeting the glutaminase (GLS)-mediated glutamine metabolism, European Review for Medical and Pharmacological Sciences, 2017, 21: 3282-3289
Xu, Feng et al. (2017) Alpha-ketoglutarate protects Streptomyces coelicolor from visible light-induced phototoxicity, Biochemistry and Biophysics Reports. 2017 Mar;9:22-28
Shao et al.,Inhibition of Polo-like Kinase 1 (Plk1) Enhances the Antineoplastic Activity of Metformin in Prostate Cancer. J. Biol. Chem., Jan 2015; 290: 2024 - 2033.
Luo et al., Cancer Usurps Skeletal Muscle as an Energy Repository. Cancer Res., Jan 2014; 74: 330 - 340.
Hai Yan et al., Disruption of Wild-Type IDH1 Suppresses D-2-Hydroxyglutarate Production in IDH1-Mutated Gliomas. Cancer Res., Jan 2013; 73: 496 - 501
For more citations of this product click here