IDH1, Recombinant, Human, FLAG-Tag (Isocitrate Dehydrogenase [NADP] Cytoplasmic, IDH, Cytosolic NADP

IDH1 belongs to two distinct subclasses. The protein is the NADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. This protein 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. Description: Recombinant human IDH1 (aa 1-414, GenBank accesssion #NM_005896, UniProt O75874) with C-terminal FLAG tag, expressed in Sf9 insect cells via a baculovirus expression system. Molecular Weight: ~47.6kD, Specific Activity: >5600pmol/min/ug, Assay Conditions: Reaction was conducted in 50mM Tris (pH 7.4), 10mM MgCl2, 200uM DL-isocitrate, and 200uM NADP. Incubation was carried out for 30 minutes at room temperature. Absorbance was read at 340nm using Tecan Infinite 1000 (5 flash/10ms settle time). Applications: Suitable for use in the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Storage and Stability: Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.