Mouse anti human cd11b apc cy7

Rabbit anti-human SAP polyclonal antibody (pAb) (565191, Calbiochem), rabbit anti-human CRP pAb (235752, Calbiochem), rabbit anti-human C1q pAb (A0136, Dako), mouse anti-human complement component C5b-9 monoclonal antibody (mAb) (IgG2a) (011-01, Antibody Shop), mouse IgG1 isotype control (BD Biosciences), mouse IgG2a isotype control (BD Biosciences), rabbit IgG isotype control (Invitrogen), FITC-conjugated goat anti-rabbit pAb (F1262, Sigma-Aldrich), FITC-conjugated goat anti-mouse pAb (F0479, Dako), mouse anti-human CD45 FITC/CD14 PE (342408, BD Biosciences), mouse anti-human CD11b APC-Cy7 (560914, BD Biosciences), Alexa-555-conjugated goat anti-mouse IgG (A-21424, Thermo Fischer), biotinylated goat anti-rabbit IgG (BA-1000, Vector Laboratories), or biotinylated goat anti-mouse IgG (BA-9200, Vector Laboratories) are the commercial antibodies.

Staining of immune cells extracted by ED was performed by blocking the Fc receptor using FcR Blocker (Miltenyi Biotec, Bergisch Gladbach, Germany). 7AAD viability dye (eBioscience) was then added, followed by staining with mouse anti-human CD11b-APC-Cy7 (BD Biosciences), mouse anti-human CD33-FITC (BioLegend, San Diego, USA), mouse anti-human HLA-DR-PE (BD Biosciences), CD14-PE-Cy7 (eBioscience) and mouse anti-human CD15-APC (BioLegend). After incubation at 4 °C for 25 min, samples were washed twice with PBS and the pellets were resuspended in 300 µl flow cytometry staining buffer (eBioscience) and analyzed using BD FACSCanto II flow cytometer. Some tumor-infiltrating immune cells were also stained for ARG1 expression, as described above, with the addition of Fixable Viability Dye eFluor^® 780 (FVD780; eBioscience) to gate live cells. Flow cytometric data were analyzed using BD FACSuite software (BD Biosciences).