Recombinant IDH1 protein

Catalog No: 31610 Format: 100 µg $410 Buy
Expressed In: E. coli Protein Species: Human

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Contents

Recombinant IDH1 protein is supplied in 25 mM Tris pH 8.0, 300 mM NaCl, 5% glycerol. Please refer to product insert upon arrival for lot-specific concentration.

Background

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.

Application Notes

This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Protein Details

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

 
Recombinant IDH1 protein, SDS-PAGE gel.

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.

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.

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.

Storage

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

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.