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.
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 80 µg/ml FLAG peptide, 0.04% Tween-20 and 20% glycerol.
MW
48 kDa
Amino Acids
1–414(end)
Specific Activity
≥ 53 pmol/min/µg
Genbank #
NM_005896
UniProt #
O75874
Tag(s)
C-terminal FLAG-tag
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.
References
1. Dang, L., et al. (2010). Trends Mol. Med. 16(9):387-97. 2. Paschka, P., et al. (2010). J. Clin. Oncol. 28(22):3636-43.
IDH reductive activity was measured in 200 µl 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.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
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