Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids,
Assay type:Enzyme activity,
Sensitivity:> 0.01 mU/well,
Assay time:45m,
Assay Platform:Microplate reader
Product details:
Abcam's Isocitrate Dehydrogenase Assay Kit (Colorimetric) provides a convenient tool for sensitive detection of NAD(+) / NADP(+)-dependent or both IDHs in a variety of samples. The IDHs utilize isocitrate as a specific substrate leading to a proportional color development and can be easily quantified colorimetrically (λ = 450 nm) with detection sensitivity as low as 0.01 mU.
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Isocitrate dehydrogenase assay protocol summary:
- add samples and standards to wells
- incubate for 3 min and analyze with microplate reader
- incubate for 30 min - 2 hr and analyze again
This product is manufactured by BioVision, an Abcam company and was previously called K756 Isocitrate Dehydrogenase Activity Colorimetric Assay Kit. K756-100 is the same size as the 100 test size of ab102528.
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.
Isocitrate Dehydrogenase often referred to as IDH includes multiple isoforms such as IDH1 and IDH2. These enzymes catalyze the oxidative decarboxylation of isocitrate to alpha-ketoglutarate (α-KG) producing NADPH from NADP+ in the process. IDH1 weighs approximately 46 kDa and is located in the cytoplasm and peroxisomes while IDH2 operates in the mitochondria. Both isoforms of IDH are essential in cellular metabolism contributing to the citric acid cycle.
Biological function summary
Isocitrate conversion catalyzed by IDH maintains the balance of cellular NADP+/NADPH which is important for protecting cells from oxidative stress. NADPH acts as a reducing agent in several anabolic reactions such as fatty acid and nucleotide synthesis. IDH does not function as part of a large complex but its action significantly impacts redox homeostasis within cells.
Pathways
IDH plays a critical role in the citric acid cycle converting isocitrate to α-KG an important intermediate. This reaction connects to multiple metabolic pathways that influence energy production. Furthermore the production of α-KG links IDH activity with related proteins like α-Ketoglutarate dehydrogenase and glutamate dehydrogenase which further process metabolic intermediates in the cycle.
Mutations in the IDH1 and IDH2 genes link to various cancers such as glioma and acute myeloid leukemia (AML). These mutations often cause a neomorphic enzyme activity producing 2-hydroxyglutarate instead of α-KG which disrupts cellular metabolism and contributes to oncogenesis. The abnormal 2-hydroxyglutarate production interrelates with proteins involved in epigenetic regulation affecting DNA and histone methylation states which are frequently altered in cancerous cells.
