Brilliant violet 510 anti mouse human cd11b

For flow cytometry analysis of in vitro mouse immune cells, the mouse T cells were incubated with 1× Golgiplug (BD Biosciences) for 6 hours. Immune cells were stained with surface antibodies in PBS for 30 minutes as described previously (51 (link)). Fixation and permeabilization processes were carried out with fixation buffer (BD Biosciences) according to the manufacturer’s protocols. The immune cells were then stained with antibodies in PBS to detect proteins. The following antibodies were used for staining: Brilliant Violet 510 anti-mouse/human CD11b (BioLegend, catalog 101245), FITC anti-mouse F4/80 antibody (BioLegend, catalog 123107), PerCP/Cyanine5.5 anti-mouse CD4 antibody (BioLegend, catalog 100540), APC anti-mouse CD206 antibody (BioLegend, catalog 141708), Brilliant Violet 421 anti-mouse CD86 antibody (BioLegend, catalog 105032), and PE anti-mouse FoxP3 antibody (Elabscience, catalog E-AB-F1238D). The flow cytometry was run using FACSCelesta flow cytometer (BD Biosciences), and the results were analyzed with FlowJo V10.7.1.

Tumors were enzymatically digested to single cell suspensions and filtered twice through 70 μm filters. Cells were then resuspended in staining buffer (DPBS containing 3% FBS) and stained with the indicated antibodies: APC/Cyanine7 anti-mouse CD45 (Cat. 147717, BioLegend), PE anti-mouse CD3 (Cat. 100205, BioLegend), APC anti-mouse CD8a (Cat. 100711, BioLegend), PerCP/Cyanine5.5 anti-human/mouse Granzyme B (Cat. 372211, BioLegend), VioletFluor™ 450 Anti-Mouse IFN gamma (Cat 75-7311-U025, Tonbo Bioscience), Brilliant Violet 510™ anti-mouse/human CD11b (Cat. 101245, BioLegend), PE anti-mouse Ly-6C (Cat. 128007, BioLegend), FITC anti-mouse Ly-6G (Cat.127605, BioLegend), PerCP/Cyanine5.5 anti-mouse F4/80 (Cat. 123127, BioLegend). Flow cytometry was performed on BD FACS Symphony machines. Flow analyses were performed using FlowJo 10.6.1 software (TreeStar). All flow antibodies were used at a 1:200 dilution.