IMP-6610

IDH1

Recombinant IDH1 protein was expressed in E. coli cells as the full length protein (accession number AAH93020.1) with a C-terminal 6xHis Tag. The molecular weight of the protein is 47.7 kDa.

IDH1 (Isocitrate Dehydrogenase (NADP(+)) 1, Cytosolic, also known as HEL-216, HEL-S-26, IDCD, IDH, IDP, IDPC, PICD) is a member of isocitrate dehydrogenases, which catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. Isocitrate dehydrogenases belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer. IDH is the NADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence. The presence of this enzyme in peroxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2, 4-dienoyl-CoAs to 3-enoyl-CoAs, as well as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production.

E. coli

Human

His

Recombinant IDH1 protein is supplied at a concentration of 0.8 μg/μL in 25 mM Tris pH 8.0, 300 mM NaCl, 5% glycerol.

47.7 kDa

>90%

Enzyme kinetics, screening inhibitors, and selectivity profiling

Recombinant proteins in solution are temperature sensitive and must be stored at -80 centigrade to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Recombinant IDH1 protein, SDS-PAGE gel. Recombinant IDH1 protein was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 47.7 kDa Purity: >90%

Recombinant IDH1 protein activity assay. 200 μM DL-isocitric acid trisodium and 200 μM NADP+ were incubated with IDH1 protein in reaction buffer including 50 mM Tris-HCl pH 7.4, 100 mM MgCl2 for 10 minutes at room temperature. The generation of product NADPH was monitored by measuring absorbance at 340 nm.

Recombinant IDH1 protein activity assay. 200 uM DL-isocitric acid trisodium and 200 uM NADP+ were incubated with IDH1 protein in reaction buffer including 50 mM Tris-HCl pH 7.4, 100 mM MgCl2 for 10 minutes at room temperature. The generation of product NADPH was monitored by measuring absorbance at 340 nm.