Alpha Ketoglutarate (alpha KG) Assay Kit provides a simple, sensitive and rapid means for quantifying alpha KG.
Colorimetric/Fluorometric
Tissue, Suspension cells, Adherent cells
Quantitative
0.01 - 10 nmol/well
30m
= 0.01 nmol/well
Alpha Ketoglutarate (alpha KG) Assay Kit provides a simple, sensitive and rapid means for quantifying alpha KG.
Colorimetric/Fluorometric
Tissue, Suspension cells, Adherent cells
Quantitative
0.01 - 10 nmol/well
30m
Microplate reader
= 0.01 nmol/well
Blue Ice
-20°C
-20°C
-20°C
Alpha Ketoglutarate (alpha KG) Assay Kit provides a simple, sensitive and rapid means for quantifying alpha KG.
In the alpha KG assay protocol, alpha-ketoglutarate is transaminated with the generation of pyruvate, which is used to convert a nearly colorless probe to a colored (λmax = 570 nm) and fluorescent (Ex/Em = 535/587 nm) product.
Alpha Ketoglutarate assay protocol summary:
- add reaction mix to standard and sample wells
- incubate for 30 min
- analyze with a microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K677 Alpha-Ketoglutarate Colorimetric/Fluorometric Assay Kit. K677-100 is the same size as the 100 test size of ab83431.
This supplementary information is collated from multiple sources and compiled automatically.
Alpha-ketoglutarate sometimes called alpha-ketoglutaric acid or A-KG is an important intermediate in several metabolic processes. It plays a mechanical role in the Krebs cycle also known as the citric acid cycle where it functions to transfer energy through oxidative reactions. A-KG is a five-carbon molecule with a mass of approximately 146.11 g/mol and is primarily expressed in the mitochondria of cells particularly within tissues with high metabolic activity such as liver and muscle.
Alpha-ketoglutarate supports amino acid metabolism and the regulation of gene expression. It acts as an essential nitrogen transporter in biochemical pathways. A-KG serves as a co-substrate for several dioxygenases including prolyl hydroxylases which are enzymes connected to collagen synthesis and the hypoxia-inducible factor pathway. Additionally it is involved in the formation of neurotransmitters and the detoxification of ammonia in animals.
Alpha-ketoglutarate is deeply integrated within the tricarboxylic acid cycle and the gamma-aminobutyric acid (GABA) shunt. Within the TCA cycle A-KG forms important energy carriers like NADH and FADH2 impacting energy production. In the GABA shunt it closely associates with glutamate dehydrogenase which transforms glutamate emphasizing its role in nitrogen and energy balance. A-KG interacts with proteins like succinyl-CoA synthetase and is linked with enzymes such as isocitrate dehydrogenase.
Alterations in alpha-ketoglutarate levels have been linked to metabolic disorders like hyperammonemia and certain types of cancer. In hyperammonemia improper detoxification processes through the urea cycle can lead to elevated ammonia levels a problem in which alpha-ketoglutarate plays a detoxifying role. Oncogenic mutations in isocitrate dehydrogenase can lead to abnormal A-KG levels contributing to cancer progression by affecting cellular metabolism and gene expression.
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Standard curve: mean of duplicates (+/- SD) with background reads subtracted
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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