Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 998736
View all Indoleamine 2,3-dioxygenase/IDO Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived human Indoleamine 2,3-dioxygenase/IDO
Ala2-Gly403
Accession # P14902

Specificity

Detects human Indoleamine 2,3-dioxygenase/IDO in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

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

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

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

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 antibody in Human Tonsil Tissue by Immunohistochemistry (IHC-P).

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 antibody in Human Monocytes antibody by Flow Cytometry.

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

Flow Cytometry Panel Builder

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Flow Cytometry

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

  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.

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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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

For research use only

Citations for Human Indoleamine 2,3‑dioxygenase/IDO Antibody

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Protocols

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