Protocol SDS

Application

• For quantitative colorimetric kinetic determination of isocitrate dehydrogenase activity and evaluation of drug effects on its metabolism.

Key Features

• 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-incubate-measure” type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.

Method

• OD565nm

Samples

• Plasma, serum, tissue and culture media, etc

Species

• All

Procedure

• 30 min

Size

• 100 tests

Detection Limit

• 0.1 U/L

Shelf Life

• 6 months

More Details

ISOCITRATE DEHYDROGENASE (IDH) is an enzyme that catalyzes the interconversion of isocitrate and a-ketoglutarate. There are three IDH isoforms: IDH3 uses the cofactor NAD⁺ and catalyzes the third step in the citric acid cycle, while IDH1 and IDH2 use the cofactor NADP⁺ and catalyze the same reaction outside the citric acid cycle. This kit measures the activity of the NADP⁺ isoforms. Mutations in IDH1 and IDH2 have been linked with various brain tumors and acute myeloid leukemia. BioAssay Systems’ non-radioactive, colorimetric IDH assay is based on the reduction of the tetrazolium salt MTT in an 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.