Cd45 bv421 anti mouse antibody

Filtered cells were pelleted down with gentle centrifugation and incubated with 50 μl of Fc blocker anti-mouse CD16/32 (Cat:101321, Biolegend) on ice for 10 min. Then 50 μl of CD11b-PE Rat anti-mouse antibody (Cat: 553311, BD bioscience) and CD45 BV421 anti-mouse antibody (Cat: 563890, BD bioscience) were mixed with the cells and incubated for 30 min on ice. Following incubation, cells were washed and resuspended in 200 μl of FACS buffer. Cells were filtered through a 30 μm filter before FACS sorting (BD FACS Aria Fusion) and collecting approximately 20,000-100,000 CD11b⁺; CD45^med cells in FACS buffer. Unstained and single-stained cells were used as controls for gating the cells in FACS. Sorted cells were centrifuged at 6800 g for 15 min and the cell pellets were snap frozen and stored at −80 °C until RNA extraction.

Filtered cells were pelleted down with gentle centrifugation and incubated with 50μl of Fc blocker anti-mouse CD16/32 (Cat: 101321, Biolegend) on ice for 10min. Then 50μl of CD11b-PE Rat anti-mouse antibody (Cat: 553311, BD Bioscience) and CD45 BV421 anti-mouse antibody (Cat: 563890, BD Bioscience) were mixed with the cells and incubated for 30min on ice. Following incubation, cells were washed and resuspended in 200μl of FACS buffer. Cells were filtered through a 30μm filter before FACS sorting (BD FACS Aria Fusion) and collecting approximately 20,000–100,000 CD11b⁺; CD45^med cells in FACS buffer. Unstained and single-stained cells were used as controls for gating the cells in FACS. Sorted cells were centrifuged at 6800g for 15min and the cell pellets were snap frozen and stored at -80°C until RNA extraction.