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AB272530

Isocitrate Dehydrogenase Assay Kit

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This kit (ab272530) measures the activity of the NADP+ isoforms.

View Alternative Names

PICD, IDH1, Isocitrate dehydrogenase [NADP] cytoplasmic, IDH, Cytosolic NADP-isocitrate dehydrogenase, IDPc, NADP(+)-specific ICDH, Oxalosuccinate decarboxylase

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Enzyme activity assay - Isocitrate Dehydrogenase Assay Kit (AB272530)
  • Enzyme activity assay

Supplier Data

Enzyme activity assay - Isocitrate Dehydrogenase Assay Kit (AB272530)

Isocitrate Dehydrogenase Assay Kit Titration Curves.

Typical standard curve – data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.

Example of Isocitrate Dehydrogenase titration curves.

OD565 readings (left) and calculated activity (right).

Key facts

Sample types

Plasma, Serum, Tissue Culture Media

Range

0.1 - 100 U/L

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Reactivity data

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Product details

This kit (ab272530) measures the activity of the NADP+ isoforms. Mutations in IDH1 and IDH2 have been linked with various brain tumors and acute myeloid leukemia. Our non-radioactive, colorimetric IDH assay is based on the reduction of the tetrazolium salt MTT in a NADPH-coupled enzymatic reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The increase in absorbance at 565 nm is directly proportional to the enzyme activity.

Fast and sensitive. Linear detection range (20 μL sample): 0.1 to 100 U/L for 30 min reaction.

Convenient and high-throughput. Homogeneous "mix-incubatemeasure" type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.

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What's included?

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Properties and storage information

Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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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.
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Product protocols

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Target data

Catalyzes the NADP(+)-dependent oxidative decarboxylation of isocitrate (D-threo-isocitrate) to 2-ketoglutarate (2-oxoglutarate), which is required by other enzymes such as the phytanoyl-CoA dioxygenase (PubMed : 10521434, PubMed : 19935646). Plays a critical role in the generation of NADPH, an important cofactor in many biosynthesis pathways (PubMed : 10521434). May act as a corneal epithelial crystallin and may be involved in maintaining corneal epithelial transparency (By similarity).
See full target information IDH1
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websiteProtocolBooklet
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