Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Tissue, Suspension cells, Adherent cells,
Assay type:Quantitative,
Sensitivity:= 0.01 nmol/well,
Range:0.01 - 10 nmol/well,
Assay time:30m,
Assay Platform:Microplate reader
Product details:
Alpha Ketoglutarate (alpha KG) Assay Kit provides a simple, sensitive and rapid means for quantifying alpha KG.
Alpha ketoglutarate assay principle
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
Other Notes
This product was previously called K677 Biovision Alpha-Ketoglutarate Colorimetric/Fluorometric Assay Kit. Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
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.
Biological function summary
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.
Pathways
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.
