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.

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 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.

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

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.

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

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Intracellular Staining by Flow Cytometry

2.5 µg/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), and Human Monocytes selected from PBMC using MagCellect Human CD14+ Cell Isolation Kit (Catalog # MAGH105) and cultured with Recombinant Human MCSF (Catalog # 216-MC), Recombinant Human IFNg (Catalog # 285-IF) and LPS, were fixed with paraformaldehyde and permeabilized with saponin

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

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 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

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

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Indoleamine 2,3-dioxygenase/IDO Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB6030

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

View all Indoleamine 2,3-dioxygenase/IDO Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

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

Indoleamine 2,3-dioxygenase/IDO in A431 Human Cell Line.