Indoleamine 2,3-dioxygenase/IDO in A431 Human Cell Line.
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 counterstained 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 IgG1 Isotype 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.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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).
Lewis-Ballester, A. et al. (2009) Proc. Natl. Acad. Sci. USA. 106:17371.
Costantino, G. (2009) Expert Opin. Ther. Targets 13:247.
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