武汉云克隆科技股份有限公司

Isocitrate Dehydrogenase 2 (IDH2) catalyzes the oxidative decarboxylation of isocitrate to alpha-ketoglutarate. There are two subclasses in the IDH family, one of them utilizing NADP  as the electron acceptor and the other using NAD . The protein encoded by this gene is the NADP dependent isocitrate dehydrogenase found in the mitochondrial. The activity of recombinant human IDH2 was measured by the ability to oxidatively decarboxylate isocitrate to 2-oxoglutarate. The reaction was performed in 25 mM Tris, 0.5 mM MnCl2, 5 mM DTT, pH 7.5 (Assay Buffer), initiated by addition 50 μL of various concentrations of IDH2 (diluted by Assay Buffer) to 50 µL of substrate mixture consisted of 1 mM NADP and 2 mM isocitric acid. The final well serves as a negative control with no IDH2, replaced with 50μl assay buffer. Incubated at 25℃ for 5min, then read at a wavelength of 340 nm. The specific activity of recombinant human IDH2 is >200 pmol/min/µg.