Human Isocitrate Dehydrogenase (IDH1), also known as oxalosuccinate decarboxylase, GenBank Accession No. NM_005896, full length (a.a. 1-414(end)) with Arg to His mutation on a.a. 132 and C-terminal FLAG-tag, MW= 48 kDa, expressed in Sf9 cells via a baculovirus expression system.
Specific activity: >= 53 pmol/min/ug
Background
Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). IDH1 homodimer is an 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, as well as in peroxisomal reactions that consume 2-oxoglutarate. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production.
Immunogen Region
1-414(end)
Tag
C-terminal FLAG-tag
Formulation
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 80 ug/ml FLAG peptide, 0.04% Tween-20 and 20% glycerol.
Species
Human
Application
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling
Notes
IDH reductive activity was measured in 200 ul reaction containing 25 mM Tris (pH 7.4), 150 mM NaCl, 10 mM MgCl2, 0.03% BSA, 1 mM alpha-Ketoglutarate, 10 M NADPH and IDH. Depletion of NADPH was monitored continuously at Abs340 nm for 20 min. Molar extinction coefficient of NADPH is 6,200 M-1cm-1.