Isocitrate Dehydrogenase BioAssay™ Kit (IDH)

Isocitrate Dehydrogenase (IDH) is an enzyme which 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. The 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 565nm. The increase in absorbance at 565nm is directly proportional to the enzyme activity.

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

Applications:
IDH activity determination in biological samples (e.g. plasma, serum, tissue and culture media.)

Range:
0.1-100U/L for 30 min reaction.

Detection limit:
0.1U/L

Kit Components:
Assay Buffer, 1x10ml
NADP/MTT, 1x1ml
Substrate, 1x1ml
Diaphorase, 1x120ul
Calibrator, 1x10ml

Storage and Stability:
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.