||Expression Host:||Human 293HEK cells
|Expression cDNA Clone and Sequence
Recombinant protein was produced with TrueORF clone, RC206592. Click on the TrueORF clone link to view cDNA and protein sequences.|
||Predicted MW:||45.1 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.
||The specific activity of IDOI was determined by monitoring kynurenine formation from N-formylkynurenine based on the absorbance at 492nm. The N-formylkynurenine was produced from a conversion of tryptophan with IDO1. The reaction was carried out at 25°C for 15min in the buffer containing 100mM PBS, pH6.5, 40mM ascorbic acid, 450 units catalase, 20µM methylene blue, and 800µM L-tryptophan as the substrate. The reaction was terminated by adding 50ul of 30% (w/v) trichloroacetic acid. The sample was further incubated for 30min at 60°C and centrifuged at 12000 rpm for 15 min. The supernatant was used to mix with an equal volume of Ehrlich’s reagent (2% p-dimethylaminobenzaldehyde in glacial acetic acid) to measure the absorbance at 492 nm after 10min incubation.
||Tryptophan metabolismMetabolic pathways