Percp cyanine5.5 anti mouse human cd11b

After myelin removal of brain homogenates, the cell suspensions were stained with the following antibodies: PE anti‐mouse Ly‐6C (BioLegend, 128008, clone HK1.4, 1:200), PerCP/Cyanine5.5 anti‐mouse/human CD11b (BioLegend, 101228, clone M1/70, 1:100), APC anti‐mouse F4/80 (BioLegend, 123116, clone BM8, 1:100), PE/Cyanine7 anti‐mouse CD45 (BioLegend, 103114, clone 30‐F11, 1:100), and Live/Dead Fixable Yellow Dead Cell Stain Kit (Invitrogen, L34959, 1.500). The CD11b⁺CD45⁺Ly6C⁻F4/80^int cells were sorted and collected using a SONY SH800 Cell Sorter. For magnetic‐activated cell sorting of splenocytes, the spleens were mashed, followed by red blood cells removal using ACK lysis buffer (Gibco, A1049201). The cell suspension was first incubated with anti‐CD11b MicroBeads (MiltenyiBiotec, 130‐049‐601), and the on‐column CD11b⁺ cells were collected as the myeloid cells. The uncollected non‐myeloid (CD11b⁻) cells were processed with the Pan T Cell Isolation Kit (MiltenyiBiotec, 130‐095‐130) following the manufacturer’s protocol, and the untouched T cells were collected, whereas the on‐column cells were flushed and collected as the B cells.