Isocitrate Dehydrogenase Assay Kit (ab272530)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Sample type: Plasma, Serum, Tissue Culture Media
Overview
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Product name
Isocitrate Dehydrogenase Assay Kit
See all Isocitrate Dehydrogenase kits -
Detection method
Colorimetric -
Sample type
Serum, Plasma, Tissue Culture Media -
Assay type
Quantitative -
Range
0.1 - 100 -
Product overview
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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Tested applications
Suitable for: Functional Studiesmore details
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Assay Buffer 1 x 10ml Calibrator 1 x 1.5ml Diaphorase 1 x 120µl NADP/MTT 1 x 1ml Substrate 1 x 1ml -
Research areas
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Relevance
Isocitrate dehydrogenase (IDH; EC 1.1.1.41, NAD+) is an enzyme that participates in the citric acid cycle. These IDH3 isoforms catalyze the oxidative decarboxylation of isocitrate, producing alpha-ketoglutarate and CO2 while converting NAD+ to NADH. This is a two-step process, which involves oxidation of isocitrate to oxalosuccinate, followed by the decarboxylation of the beta-carboxyl group to form the ketone, alpha-ketoglutarate. Other isoforms (EC 1.1.1.42, NADP+) catalyze the same reaction, but unrelated to the citric acid cycle. It is carried out in the mitochondrion (IDH2) as well as in the cytosol and peroxisome (IDH1) and use NADP+ as a cofactor instead of NAD+. -
Cellular localization
Cytoplasm. Peroxisome -
Alternative names
- Cytosolic NADP isocitrate dehydrogenase
- EC=1.1.1.42
- IDH
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Images
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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).