Table 1.
Surface markers and antibody combinations for determining immune cells from the lungs and the brain following tMCAO
| Antibody | Clone | Immune cell type | Population | Surface marker expression |
|---|---|---|---|---|
| CD45‐FITC | 30‐F11 | Alveolar macrophages | L1 | CD45+ Siglec F+ CD11b− |
| Siglec F‐PE | E50‐2440 | Eosinophils | L2 | CD45+ Siglec F+ CD11b+ |
| CD11c‐Percp/Cy5.5 | N418 | CD103+ DCs | L3 | CD45+ Siglec F− CD11b− CD103+ CD11c+ MHC II+ |
| CD11b‐PE/Cy7 | M1/70 | CD11b+ DCs | L4 | CD45+ Siglec F− CD11b hi CD103− CD64− MHC II+ |
| CD64‐APC | X54‐5/7.1 | Interstitial macrophages | L5 | CD45+ Siglec F− CD11b hi CD103− CD64+ MHC II+ |
| Live/dead‐APC/Cy7 | ||||
| CD103‐BV421 | 2E7 | |||
| MHC II‐BV510 | M5/114.15.2 | |||
| CD45‐FITC | 30‐F11 | Monocytes/moDCs | L8 | CD45+ CD11b hi Ly6C hi/int CCR2+/− Ly6G− |
| Ly6C‐PE | HK1.4 | Neutrophils | L9 | CD45+ CD11b hi Ly6C int CCR2− Ly6G+ |
| CD11b‐PE/Cy7 | M1/70 | |||
| Live/dead‐APC/Cy7 | ||||
| CCR2‐BV421 | SA203G11 | |||
| Ly6G‐BV510 | 1A8 | |||
| CD45‐FITC | 30‐F11 | Plasmacytoid DCs | L6 | CD45+ B220+ CD11c+ |
| CD8‐PE | 53‐6.7 | B cells | L7 | CD45+ B220+ CD11c− |
| NK1.1‐Percp/Cy5.5 | PK136 | CD4+ T cells | L10 | CD45+ B220− CD11c− CD4+ CD8− |
| CD11c‐PE/Cy7 | N418 | CD8+ T cells | L11 | CD45+ B220− CD11c− CD4− CD8+ |
| B220‐APC | RA3‐6B2 | NK cells | L12 | CD45+ B220− CD11c− CD4− CD8− NK1.1+ TCRβ− |
| Live/dead‐APC/Cy7 | NKT cells | L13 | CD45+ B220− CD11c− CD4− CD8− NK1.1+ TCRβ+ | |
| CD4‐BV421 | GK1.5 | |||
| TCRβ‐BV510 | H57‐597 |
Abbreviations: FITC, fluorescein isothiocyanate; moDC, monocyte‐derived dendritic cell; NK, natural killer; NKT, natural killer T; tMCAO,transient middle cerebral artery occlusion.