||Expression Host:||HEK293 cells
|Expression cDNA Clone or AA Sequence
Recombinant protein was produced with TrueORF clone, RC202777
. Click on the TrueORF clone link to view cDNA and protein sequences.
||Predicted MW:||47.7 kDa|
|Purity:||> 80% as determined by SDS-PAGE and Coomassie blue staining|
|Concentration:||>50 ug/mL as determined by microplate BCA method|
|Buffer:||25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol.|
||Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
||The specific activity of TDO2 was determined by monitoring Kynurenine formation from the N-formylkynurenine based on the absorbance at 492nm. The N-formylkynurenine was produced from a conversion of tryptophan with TDO2. The reactions were carried out at 37°C for 40min in 100ul of the reaction volume containing 200mM PBS, pH7.5, 1mM ascorbic acid, and 1.25mM L-tryptophan as the substrate with various amounts of TDO2. 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-dimethylaminobenza-ldehyde in glacial acetic acid) to measure the absorbance at 492 nm after 10min incubation
||Tryptophan metabolismMetabolic pathways