||Expression Host:||Human 293HEK cells
|Expression cDNA Clone and Sequence
Recombinant protein was produced with TrueORF clone, RC210582. Click on the TrueORF clone link to view cDNA and protein sequences.|
||Predicted MW:||46.5 kDa|
|Purity:||> 80% as determined by SDS-PAGE and Coomassie blue staining|
|Concentration:||>50 ug/mL as determined by microplate BCA method|
|Buffer and Storage:||25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol. Store at -80C. Avoid repeated freeze-thaw cycles. Stable for at least 3 months from receipt of products under proper storage and handling conditions.|
||Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
||Enzymatic activities were determined by monitoring NADPH formation based on the absorbance at 345nm. The reaction was carried out at 37? for 10 minutes in the presence of isocitrate as a substrate and NADP as a cofactor. The data which presented a good linear relation on the curve was used to calculate the specific activity, and one unit is defined as converting 1.0 umole of NADP to NADPH per min at 37?. In summary, the wildtype IDH1 produced from HEK293 cells and insect cells are active while the R132H mutant or the WT/R132H heterodimers are inactive.
||Citrate cycle (TCA cycle)Glutathione metabolismMetabolic pathways