Human Indoleamine 2,3‑dioxygenase/IDO PE‑conjugated Antibody

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 (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 PE-conjugated Monoclonal Antibody (Catalog # IC6030P) or (B) Mouse IgG₁ isotype control antibody (IC002P) and Mouse Anti-Human CD14 APC-conjugated Monoclonal Antibody (FAB3832A). 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 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 PE-conjugated Monoclonal Antibody (Catalog # IC6030P) or (B) Mouse IgG₁Phycoerythrin Isotype Control (Catalog # IC002P). 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.

Detection of Human Indoleamine 2,3-dioxygenase/IDO by Flow Cytometry

SLE sera increase IDO expression in a type I IFN-dependent manner.Endothelial cells (WISH cells) were incubated with A) recombinant IFN-alpha with or without pre-incubation with an IFN alpha/beta receptor blocking antibody (IFNAR ab) and IDO expression analyzed in permeabilized cells by flow cytometry. B) WISH cells were incubated with serum from SLE patients or healthy individuals with or without pre-incubation with an IFNAR ab. The results are presented as percentage increased IDO expression in median fluorescence intensity (MFI) as compared to non-stimulated cells. C) Patients with ongoing type I IFN activity (SLE IFN+) had increased IDO activity as compared to patients with no type I IFN activity (SLE IFN-) and healthy volunteers (HV). D) Patients with ongoing type I IFN activity had decreased serotonin levels as compared to patients with no type I IFN activity and healthy volunteers. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0125109), licensed under a CC-BY license. Not internally tested by R&D Systems.