Human Monocytes were selected from PBMC using MagCellect Human CD14+ Cell Isolation Kit (MAGH105) and cultured overnight with Recombinant Human MCSF (50 ng/mL; 216-MC), Recombinant Human IFN gamma (50 ng/mL; 285-IF) and 50 ng/mL LPS and stained with (A) Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Monoclonal Antibody (Catalog # IC6030A) or (B) Mouse IgG₁ isotype control antibody (IC002A) and Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (FAB3832P). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of Indoleamine 2,3‑dioxygenase/IDO in Human MDSCs by Flow Cytometry.

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 PE-conjugated Monoclonal Antibody (Catalog # FAB1137P) and either (A) Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Monoclonal Antibody (Catalog # IC6030A) or (B) Mouse IgG₁Allophycocyanin Isotype Control (Catalog # IC002A). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Applications for Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Antibody

Application

Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/10⁶ cells
Sample: Human myeloid-derived suppressor cells (MDSCs) treated with Recombinant Human IL‑6 (Catalog # 206-IL) and Recombinant Human GM‑CSF (Catalog # 215-GM) were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005) and Human Monocytes selected from PBMC using MagCellect Human CD14+ Cell Isolation Kit (Catalog # MAGH105) and cultured overnight with Recombinant Human MCSF (50 ng/mL, Catalog # 216-MC), Recombinant Human IFN gamma (50 ng/mL, Catalog # 285-IF) and 50 ng/mL LPS

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Protect from light. Do not freeze.

12 months from date of receipt, 2 to 8 °C as supplied.

Background: Indoleamine 2,3-dioxygenase/IDO

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).

References

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

Alternate Names

3dioxygenase, IDO, IDO1, INDO, Indoleamine 2

Entrez Gene IDs

3620 (Human); 15930 (Mouse)

Gene Symbol

IDO1

UniProt

P14902

Additional Indoleamine 2,3-dioxygenase/IDO Products

Product Documents for Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Antibody

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Product Specific Notices for Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Antibody

For research use only

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Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Antibody

R&D Systems, part of Bio-Techne | Catalog # IC6030A

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Scientific Data Images for Human Indoleamine 2,3-dioxygenase/IDO APC-conjugated Antibody

Detection of Indoleamine 2,3‑dioxygenase/IDO in Human Monocytes by Flow Cytometry.