Anti foxp3 antibody clone fjk 16s

Immunohistochemistry was performed on 5‐μm‐thick frozen sections as previously described 23 using an anti‐Foxp3 antibody (clone FJK‐16s, eBioscience).

Preparation of single-cell suspension of mouse tumor tissues by mechanical grinding. All single-cell suspensions were incubated with rat anti-mouse CD16/CD32 blocking antibody (4 μg/ml) for 15 min after thorough filtration and precipitation, stained with fluorescein-conjugated antibodies, multiple washed with PBS, and resuspended in 7-AAD (exclude non-viable cells). For anti-mouse CD4 (Clone RM4-5, BD Biosciences), anti-mouse CD8 (clone 53-6.7, eBioscience), anti-mouse CD45 (clone 30-F11, Biolegend), anti-mouse CD25 (PC61.5, eBioscience), anti-mouse PD-L1 (clone 10 F.9G2, BioLegend) and anti-mouse CD31 (clone MEC 13.3, BD Biosciences) staining, after incubation for 1 h, cells were washed with PBS for three times (1500 rpm, 5 min each), then detected by flow cytometry (BD FACSCanto II). For FoxP3 staining, after incubation, cells were washed and fixed with 1 ml of fixation & permeabilization solution (BD Biosciences) for 30–60 min, and washed twice with Perm Wash (BD Biosciences). Intracellular staining with anti-FoxP3 antibody (clone FJK-16s, eBioscience) was performed for 1 h. The next steps are the same as described above.