Western blot shows lysates of human placenta tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO Monoclonal Antibody (Catalog # MAB60302) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Indoleamine 2,3-dioxygenase/IDO at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Indoleamine 2,3-dioxygenase/IDO in Human Tonsil Tissue.

Indoleamine 2,3-dioxygenase/IDO was detected in immersion fixed paraffin-embedded sections of human tonsil tissue using Mouse Anti-Human Indoleamine 2,3-dioxygenase/IDO Monoclonal Antibody (Catalog # MAB60302) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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 # MAB60302) 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 IgG2B Isotype Control (Catalog # MAB0041). 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.

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil tissue

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human PBMC monocytes fixed with 1% paraformaldehyde and permeabilized with saponin

Western Blot

2 µg/mL
Sample: Human placenta tissue

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. 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

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COA

SDS

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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 # MAB60302

Key Product Details

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Product Specifications

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

Detection of Human Indoleamine 2,3‑dioxygenase/IDO by Western Blot.