Active Indoleamine-2,3-Dioxygenase (IDO)

CD107B; INDO; Indoleamine-Pyrrole 2,3 Dioxygenase

ACTIVITY TEST

IDO (Indoleamine 2,3-dioxygenase 1) is a heme enzyme that catalyzes the first and rate-limiting step in tryptophan catabolism to N-formyl-kynurenine. This enzyme acts on multiple tryptophan substrates including D-tryptophan, L-tryptophan, 5-hydroxy-tryptophan, tryptamine, and serotonin. Thus, bioactivity of recombinant human IDO was measured through its ability to oxidize L-tryptophan to N-formyl-kynurenine, using Methylene Blue as indicator. The reaction was performed in 50 mM MES, pH 6.5 (Assay Buffer), initiated by addition 50 μL of various concentrations of IDO (diluted by Assay Buffer) to 50 µL substrate mixture of 800 μM L-tryptophan, 9000 units/mL catalase (RPC418Hu05), and 40 μM Methyene Blue in assay buffer with equal volume of 80 mM ascorbic acid in 0.405 M Tris, pH 8.0. The final well serves as a negative control with no IDO, replaced with 50μL assay buffer. The absorbance was read in 321 nm in kinetic mode for 5 minutes. The result indicated that recombinant human IDO can oxidize L-tryptophan, the specific activity is 10581 pmol/min/μg.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Blood CCL18 differentiates dendritic cells in tolerogenic cells able to prime regulatory T cells in healthy subjects Pubmed: 21803856
Clinical Transplantation Potential immunosuppressive function of plasma indoleamine 2,3-dioxygenase in patients with aGVHD after allo-HSCT Wiley: source
Indonesian Journal of Cancer Chemoprevention Secretion of Indoleamine 2, 3-Dioxygenase, an Immunomodulatory Substance, by Adipose-Derived Mesenchymal Stem Cell Iregway: Source
PLoS ONE CD40 Gene Silencing Reduces the Progression of Experimental Lupus Nephritis Modulating Local Milieu and Systemic Mechanisms PubMed: PMC3683035
Hepatology Human CD14+ CTLA-4+ regulatory dendritic cells suppress T-cell response by cytotoxic T-lymphocyte antigen-4-dependent IL-10 and indoleamine-2,3-dioxygenase production in hepatocellular carcinoma Pubmed: 23960017
Free Radical Biology and Medicine Indoleamine-2,3-dioxygenase elevated in tumor-initiating cells is suppressed by mitocans ScienceDirect: S0891584913006345
Frontiers in Oncology Cancer-Associated Fibroblasts from lung tumors maintain their immuno-suppressive abilities after high-dose irradiation Frontiersin:Source
Tissue Engineering Part A Immunogenicity and escape mechanisms of allogeneic tendon-derived stem cells Pubmed:24813640
Immunobiology. The expression of dendritic cell subsets in severe chronic rhinosinusitis with nasal polyps is altered Pubmed:24947894
Apoptosis Chronic hepatitis C virus infection triggers spontaneous differential expression of biosignatures associated with T cell exhaustion and apoptosis signaling in peripheral blood mononucleocytes Pubmed:25577277
Front Oncol Cancer-Associated Fibroblasts from Lung Tumors Maintain Their Immunosuppressive Abilities after High-Dose Irradiation PubMed: 26029659
Acta Universitatis Tamperensis Insights into the use of adipose stem cells for clinical cell therapy: Novel culturing conditions and characterization of multipotency and immunogenic properties Handle: 10024
PLoS One Respiratory Syncytial Virus-Infected Mesenchymal Stem Cells Regulate Immunity via Interferon Beta and Indoleamine-2, 3-Dioxygenase pubmed:27695127
Journal of Biomedical Nanotechnology Exploring Amino Acid-Capped Nanoparticles for Selective Anti-Parasitic Action and Improved Host Biocompatibility Pubmed:29883557
Biochemistry Mechanisms of Leptin and Ghrelin Action on Maturation and Functions of Dendritic Cells
American Journal of Pathology Cognitive Decline, Cerebral-Spleen Tryptophan Metabolism, Oxidative Stress, Cytokine Production, and Regulation of the Txnip Gene in 3xTg-AD Mice Pubmed: 30980800
international immunopharmacology Interaction of the immune-inflammatory and the kynurenine pathways in rats resistant to antidepressant treatment in model of depression Pubmed: 31176083
Stem Cell Reports Richard Schäfer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Kuçi, 4
International Journal of?Tryptophan?Research Altered Indoleamine 2, 3-Dioxygenase Production and Its Association to Inflammatory Cytokines in Peripheral Blood Mononuclear Cells Culture of Type 2 Diabetes?¡­ 33343199
Stem Cell Reports Richard Sch?fer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Ku?i, 4
Transl Psychiatry Associations between expression of indoleamine 2, 3-dioxygenase enzyme and inflammatory cytokines in patients with first-episode drug-naive … 34802039
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