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Recombinant IDO1 protein
| Catalog No: 81031 | Format: 100 µg | ¥3,720 | Buy |
| Catalog No: 81731 | Format: 1 mg | ¥24,000 | Buy |
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Expressed In: E. coli 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
IDO1 (Indoleamine 2,3-dioxygenase 1, also known as IDO, INDO, or IDO-1) is a heme enzyme that catalyzes the first and rate-limiting step in tryptophan catabolism to N-formyl-kynurenine. Tryptophan shortage inhibits T lymphocytes division and accumulation of tryptophan catabolites induces T-cell apoptosis and differentiation of regulatory T-cells. This enzyme is thought to play a role in a variety of pathophysiological processes such as antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Through its expression in dendritic cells, monocytes, and macrophages this enzyme modulates T-cell behavior by its peri-cellular catabolization of the essential amino acid tryptophan. IDO1 acts on multiple tryptophan substrates including D-tryptophan, L-tryptophan, 5-hydroxy-tryptophan, tryptamine, and serotonin.
Application Notes
This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Activity Assay Conditions: 400 µM L-tryptophan was incubated with different concentrations of IDO1 protein in reaction buffer including 50 mM potassium phosphate pH 6.5, 20 mM ascorbic acid, 10 µM methylene blue and 200 µg/ml catalase for 1 hr at 37°C. The total 200 µl reaction was stopped by the addition of 40 µl 30% (w/v) trichloroacetic acid and heated at 65°C for 15 min. After cooled down to 4°C, the reaction was centrifuged at 1,125 g for 10 min. 100 µl supernatant was transferred into 96-well plate and mixed with 100 µl of 2% (w/v) p-DMAB in acetic acid. The yellow pigment derived from kynurenine was measured at 490 nm.
Protein Details
Recombinant IDO1 was expressed in E. coli cells as the full length protein (accession number NP_002155.1) with an N-terminal 6×His tag. The molecular weight of the protein is 47.5 kDa.
Recombinant IDO1 protein gel
10% SDS-PAGE Coomassie staining
MW: 47.5 kDa
Purity: > 90%
Recombinant IDO1 protein activity assay
400 µM L-tryptophan was incubated with different concentrations of IDO1 protein in reaction buffer mixture. The reaction was stopped by the addition of 40 µl 30% (w/v) trichloroacetic acid and heated at 65°C for 15 min. After cooling down to 4°C, the reaction was centrifuged at 1,125 g for 10 min. 100 µl supernatant was transferred into 96-well plate and mixed with 100 µl of 2% (w/v) p-DMAB. The yellow pigment derived from kynurenine was measured at 490 nm.
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.
