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Recombinant Proteins & Enzymes

Recombinant IDO1 protein

Catalog No: 81031 Format: 100 µg $395 Buy Now
Catalog No: 81731 Format: 1 mg $2,600 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant IDO1 proteinContents

Recombinant IDO1 protein is supplied in 25 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol and 0.5 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant IDO1 proteinBackground

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. 

for Recombinant IDO1 proteinApplication 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. 

for Recombinant IDO1 proteinProtein 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 protein gel

Recombinant IDO1 protein gel
10% SDS-PAGE Coomassie staining
MW: 47.5 kDa
Purity: > 90% 

Recombinant IDO1 protein activity assay

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.

for Recombinant IDO1 proteinStorage

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.

for Recombinant IDO1 proteinGuarantee

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.

 

Technical Data Sheet

Recombinant IDO1 protein
 

Data Thumbnails

Recombinant IDO1 protein protein gel

Recombinant IDO1 protein protein gel
10% SDS-PAGE Coomassie staining
(Click to enlarge and view details).

Recombinant IDO1 protein activity assay

Recombinant IDO1 protein activity assay
(Click to enlarge and view details).