Anti cd15 pe

For flow cytometry analysis, tumor and blood samples were stained with a cocktail of monoclonal mouse anti-human conjugated antibodies (mAbs): anti-CD45-PercP (clone HI30), anti-CD3-PercP (HIT3a), anti-CD3-APC (UCHT1), anti-CD19-PE (HIB19), anti-CD15-PE (HI98), anti-CD161-FITC (HP-3G10), anti-CD4-FITC (OKT4), anti-CD8-PE (HIT8a), anti-HLA-DR-APC (L243), anti-CD127-PE-Cy7 (A019D5), anti-CD1c-APC-Cy7 (L161), anti-CD163-PE (GHI/61), anti-CD206-APC-Cy7 (15–2), anti-PD-1-FITC (EH12.2H7), anti-PD-L1-APC (29E2A3), anti-CTLA4-PE (L3D10), anti-CD69-APC-Cy7 (FN50), anti-Tim3-APC (F38-2E2), anti-IL-8-APC (E8N1), anti-IFN-γ-PE (4S.B3), anti-IFN-γ-APC-Cy7 (4S.B3), anti-Granzyme B-FITC (QA16A02), anti-IL-1β-FITC (JK1B-1), anti-IL-2-PE-Cy7 (MQ1-17H12), anti-IL-6-APC (MQ2-13A5), anti-IL-17-FITC (BL168), anti-IL-23/IL-12-PE (C11.5), anti-TGF-β-APC (TW4-6H10), all from Biolegend; anti-IDO-PE (eyedio) and anti-IL-10-FITC (BT-10), both from eBioscience; anti-CD25-PE (MEM-181) and anti-CD11b-FITC (LT11) from ImmunoTools.
The antibodies used for immunofluorescence were: mouse monoclonal anti-human CD8 (32-M4) from Santa Cruz Biotechnology and rabbit polyclonal anti-human HLA-DRA from Sigma Aldrich.

Cell viability was evaluated at the time of retinae cell dissociation. To ensure we obtain live cells after sorting, we labeled the cells with Zombie Aqua^TM (BioLegend, San Diego, CA, USA) a permeant dye to discriminate between live (negative for dye) and dead (positive for dye) cells. Live cells were treated with 1.0 μg of anti-CD16/CD32 per 1.0 × 10⁶ cells in 100 μL to block FcγRII/III (clone 93; BioLegend), which minimizes non-specific binding of the primary antibodies and in turn inhibits endocytosis, phagocytosis and antigen presentation due to FcγR activation. The following primary antibodies were used to detect surface antigens by incubating the cells on ice for 30 min: anti-CD90.1 PerCP-Cy5.5 (Thy1.1, clone OX-7, BioLegend, exhibits no cross-reactivity with CD90.2); anti-CD90.2 Alexa Fluor-700 (Thy1.2, clone 30-H12, BioLegend, exhibits no cross-reactivity with CD90.1); anti-CD48 PE-Cy7 (clone HM48-1, BioLegend, labels monocytes and microglia); anti-CD15 PE (clone MC-480, BioLegend, labels amacrine cells); and anti-CD57 (clone VC1.1, Sigma Aldrich, St. Louis, MO, USA also labels amacrine cells). Because the anti-CD57 antibody was unconjugated, a Brilliant violet 421-tagged secondary antibody (Life Technologies, Carlsbad, CA, USA) was used to allow for sorting.