| Species: | Human |
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. |
| Tag: | C-terminal MYC/DDK |
Predicted MW: | 45.1 kDa |
| Purity: | > 80% as determined by SDS-PAGE and Coomassie blue staining |
| Mass Spec Validation: |
This protein has been positively validated by MS/MS by Dr. Robert Moritz at The Institute of Systems Biology. |
| 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. |
| Preparation: |
Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps. |
| BioActivity: |
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. |
| Protein Families: |
Druggable Genome |
| Protein Pathways: |
Tryptophan metabolismMetabolic pathways |
