Indoleamine 2,3-dioxygenase/IDO Antibody (700838) [Unconjugated]

Indoleamine 2,3-dioxygenase/IDO was detected in immersion fixed A431 human epithelial carcinoma cells stimulated with 0.5 ng/mL of Recombinant Human IFN-gamma (Catalog # 285-IF) using Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO Monoclonal Antibody (Catalog # MAB6030) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counter-stained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Human PBMC-derived myeloid-derived suppressor cells (MDSCs) treated with 10 ng/mL Recombinant Human IL-6 (Catalog # 206-IL) and 10 ng/mL Recombinant Human GM-CSF (Catalog # 215-GM) for 7 days were stained with Mouse Anti-Human Siglec-3/CD33 APC-conjugated Monoclonal Antibody (Catalog # FAB1137A) and either (A) Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO Monoclonal Antibody (Catalog # MAB6030) or (B) Mouse IgG1Isotype Control (Catalog # MAB002) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.

Human Monocytes were selected from PBMC using MagCellect Human CD14+ Cell Isolation Kit (Catalog # MAGH105) and cultured overnight with (A) 50 ng/mL Recombinant Human MCSF (Catalog # 216-MC), 50 ng/mL Recombinant Human IFNg (Catalog # 285-IF) and 50 ng/mL LPS, or (B) Recombinant Human MCSF alone. Cells were stained with Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO Monoclonal Antibody (Catalog # MAB6030) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (Catalog # FAB3832P). Quadrant markers were set based on Mouse IgG1 Isotype Control (Catalog # MAB002). To facilitate intracellular staining, cells were fixed with 1% paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.

• Reactivity: Hu
• Applications: CyTOF-ready, Flow, ICC/IF
• Clone: 700838
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Indoleamine 2,3-dioxygenase/IDO Antibody (700838) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Indoleamine 2,3-dioxygenase/IDO
  Ala2-Gly403
  Accession # P14902
• Specificity: Detects human Indoleamine 2,3-dioxygenase/IDO in direct ELISA.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: IDO1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 8-25 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Indoleamine 2,3-dioxygenase/IDO Antibody (700838) [Unconjugated]

• 3dioxygenase
• EC 1.13.11.52
• IDO
• IDO1
• IDOIDO-1
• INDO
• INDOindole 2,3-dioxygenase
• Indoleamine 2
• indoleamine 2,3-dioxygenase 1
• Indoleamine 2,3-dioxygenase
• indoleamine-pyrrole 2,3 dioxygenase
• Indoleamine-pyrrole 2,3-dioxygenase

Background

Indoleamine 2,3-dioxygenase (IDO) is a heme-containing intracellular dioxygenase catalyzing the degradation of the essential amino acid L-tryptophan to N-formyl-kynurenine (1). This degradation is the first and rate-limiting step of the L-kynurenine pathway (2). IDO is widely expressed in dendritic cells, macrophages, microglia, eosinophils, fibroblasts, endothelial cells, and most tumor cells. In immune cells, its expression is mainly induced by cytokines such as IFN-gamma , IFN-alpha , IFN-beta , and IL‑10. IDO has an antimicrobial function due to its decreasing the availability of the essential amino acid tryptophan in inflammatory environments (3). Recent studies have demonstrated that IDO induces immunosuppression during infection, pregnancy, transplantation, autoimmunity, and neoplasia (3-5).

1. Lewis-Ballester, A. et al. (2009) Proc. Natl. Acad. Sci. USA. 106:17371.
2. Costantino, G. (2009) Expert Opin. Ther. Targets 13:247.
3. Xu, H. et al. (2008) Immunol. Lett. 121:1.
4. Lob, S. et al. (2009) Nat. Rev. Cancer 9:445.
5. Curti, A. et al. (2009) Blood 113:2394.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Indoleamine 2,3-dioxygenase/IDO Antibody (MAB6030)(3)

Publications using MAB6030

• A Gustafsson, Z Prgomet, S Jankovskaj, T Ruzgas, J Engblom, L Ohlsson, A Gjörloff W Effect of IFN-gamma on the kynurenine/tryptophan ratio in monolayer-cultured keratinocytes and a 3D reconstructed human epidermis model J. Dermatol. Sci., 2020-07-17;0(0):. 2020-07-17 [PMID: 32782183] (IHC, Human)
• Samira Mansouri, Divya S. Katikaneni, Himanshu Gogoi, Mauricio Pipkin, Tiago N. Machuca, Amir M. Emtiazjoo et al. Lung IFNAR1hi TNFR2+ cDC2 promotes lung regulatory T cells induction and maintains lung mucosal tolerance at steady state Mucosal Immunology 2020-07-01 [PMID: 31959883] (Flow Cytometry, Transgenic Mouse)
• Bonanno G, Mariotti A, Procoli A, Folgiero V, Natale D, De Rosa L, Majolino I, Novarese L, Rocci A, Gambella M, Ciciarello M, Scambia G, Palumbo A, Locatelli F, De Cristofaro R, Rutella S Indoleamine 2,3-dioxygenase 1 (IDO1) activity correlates with immune system abnormalities in multiple myeloma. J Transl Med, 2012-12-11;10(0):247. 2012-12-11 [PMID: 23232072] (Western Blot, Human)

Bioinformatics

• Gene Symbol: IDO1
• Uniprot:
  • P14902()