Allophycocyanin apc conjugated anti cd11b

The following fluorochrome-conjugated anti-mouse monoclonal antibodies for cell surface markers and intranuclear factor were purchased from eBiosciences (San Diego, California, USA): fluorescein isothiocyanate-conjugated anti-CD4 (cat no. 11-0041-85), anti-Gr-1 (cat no. 11-5931-82), anti-MHC-II (cat no. 11-5321-82); phycoerythrin-conjugated anti-CD45 (cat no. 12-0451-83), anti-CD25 (cat no. 12-0251-83); phycoerythrin cyanine7-conjugated anti-CD8 (cat no. 25-0081-82), anti-F4/80 (cat no. 25-4801-82); allophycocyanin (APC)-conjugated anti-CD11b (cat no. 17-0112-82), and anti-Foxp3 (cat no. 17-5773-82). Single-cell suspensions of splenocytes, tumor-infiltrating lymphocytes, and ascitic cells were stained on ice for 30 min with the indicated cell surface marker antibodies (dilution, 1: 200). For intranuclear Foxp3 staining, cells were fixed and permeabilized using a Cytofix/Cytoperm Kit (cat no. 00-5523-00; eBiosciences) on ice for 30 min after labeling with surface marker antibodies, followed by anti-Foxp3 mAb (dilution, 1: 50) intranuclear staining on ice for 30 min. Samples were acquired on a BD FACScanto II flow cytometer (BD Biosciences, San Jose, California, USA) and the results were analyzed using Flowjo software (TreeStar, Ashland, Oregon, USA).