Recombinant IDH2 (R172K) protein

Aliases: Isocitrate Dehydrogenase (NADP(+)) 2, Mitochondrial, NADP(+)-Specific ICDH
Catalog No: 31618 Format: 100 µg $485 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

IDH2 (Isocitrate Dehydrogenase (NADP(+)) 2, Mitochondrial, also known as MNADP-IDH, D2HGA2, IDHM, IDH, IDPM, IDC-M) is a member of isocitrate dehydrogenases, which catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. IDH2 is found in the mitochondria. It plays a role in intermediary metabolism and energy production. This protein may tightly associate or interact with the pyruvate dehydrogenase complex.

Mutations in IDH1 and IDH2 frequently occur in grade II-III gliomas, secondary glioblastomas and acute myeloid leukemias (AML). To date, mutations in at least four active site arginine residues IDH1 R100, IDH1 R132, IDH2 R140, and IDH2 R172 have been shown to result in the neomorphic production of R(-)-2-hydroxyglutarate (2HG), although these mutants lack the wild-type enzyme’s ability to convert isocitrate to α-ketoglutarate (α-KG, 2OG). 

Application Notes

This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Activity Assay Conditions: 10 µM NADPH and 1 μM α-KG were incubated with 100 nM IDH2 (R172K) protein in 200 µl reaction system containing 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 10 mM MgCl2 and 0.03% BSA (at room temperature). Depletion of NADPH was monitored continuously at Abs 340 nm for 20 min

Protein Details

Recombinant IDH2 (R172K) protein was expressed in baculovirus system as the full length protein (accession number NP_002159.2) with a point mutation Arg172Lys and a C-terminal FLAG tag. The molecular weight of the protein is 51.7 kDa.

 

Recombinant IDH2 (R172K) protein gel
12.5% SDS-PAGE Coomassie staining
MW: 51.7 kDa
Purity: > 85%

Recombinant IDH2 (R172K) protein activity assay
10 µM NADPH and 1 µM α-KG were incubated with 100 nM IDH2 (R172K) protein in 200 µl reaction system (50 mM Tris-HCl pH 7.4, 150 mM NaCl. 10 mM MgCl2 and 0.03% BSA, room temp). Depletion of NADPH was monitored continuously at Abs 340 nm for 20 min.

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.