Table 1 Antibodies applied for flow cytometry, immunohistochemistry and fluorescence stainings.
Cell population | Antibody | Isotype | Species of origin | Cat. # |
|---|---|---|---|---|
Flow cytometry: | Â | Â | Â | Â |
T-cells | TCRβ-APC | IgG2 | Hamster | 553174, BD Biosciences* |
Granulocytes | GR1-FITC | IgG2b | Rat | 553126, BD Biosciences |
TNF producing cells | TNF-PE | IgG1 | Hamster | 559503, BD Biosciences |
IL-1β producing cells | IL-1β |  | Rabbit | AAM13G, Serotec |
Microglia/macrophages | CD11b-PE/-APC/-PerCP-Cy5.5 | IgG2b | Rat | 557397/553312/550993, BD* |
Microglia/macrophages | CD45-PE/-APC/-PerCP-Cy5.5 | IgG2b | Rat | 553081/559864/550994, BD* |
Isotype/serum controls: | PE-/APC-/PerCP-Cy5.5-conjugated | IgG2b | Rat | 553989/556924/550764, BD* |
| Â | FITC-conjugated | IgG2b | Rat | 556923, BD Biosciences |
| Â | PE-conjugated | IgG1 | Hamster | 554711, BD Biosciences |
| Â | APC-conjugated | IgG2 | Hamster | 558141, BC Biosciences |
| Â | Un-conjugated | Ig fraction | Rabbit | X0903, DakoCytomation |
Secondary antibodies: | Â | Â | Â | Â |
Anti-rabbit I | Alexa Fluor647-conjugated | IgG | Goat | A21244, Invitrogen |
Anti-rabbit II | Alexa Fluor488-conjugated | IgG | Chicken | A21441, Invitrogen |
Immunohistochemistry: | Â | Â | Â | Â |
BM-derived cells | GFP (1:1000) | Â | Rabbit | ab290, Abcam |
TNF producing cells | TNF (1:50) | Â | Rat | MM-350, Endogen |
TNF producing cells | TNF (1:200) | Â | Rabbit | P-350, Endogen |
IL-1β producing cells | IL-1β (1:50) |  | Rabbit | AAM13G, Serotec |
Microglia/macrophages | CD11b (1:600) | IgG2b | Rat | MCA711, Serotec |
|  | CD45.1 Biotin-conjugated (1:80) | IgG2a,κ | Mouse | 553774, BD Biosciences |
|  | CD45.2 Biotin-conjugated (1:80) | IgG2a,κ | Mouse | 553771, BD Biosciences |
| Â | CD45 (1:80) | IgG1 | Rat | MCA1388, Serotec |
Granulocytes | Ly-6G and Ly-6C(Gr1) (1:200) | IgG2b | Rat | 01211A, BD Biosciences |
Endothelial | von Willebrandt's Factor (1:200) | Â | Rabbit | A0082, DakoCytomation |
Endothelial | PECAM-1/CD31 (1:100) | IgG2a | Rat | 557355, BD Biosciences |
Astrocytes | GFAP (1:1,200) | Â | Rabbit | Z0334, DakoCytomation |
Astrocytes | GFAP (1:50) | Â | Goat | sc-6170, Santa Cruz |
| Â | Â | Â | Â | Biotechnology |
Isotype/serum controls: | Un-conjugated | IgG2b | Rat | IG-851125, Biosite |
| Â | Un-conjugated | IgG2a | Rat | P54605M, Biosite |
| Â | Un-conjugated | Serum | Goat | X0907, DakoCytomation |
| Â | Un-conjugated | Ig fraction | Rabbit | X0903, DakoCytomation |
|  | Biotin-conjugated | IgG2a,κ | Mouse | 553455, BD Biosciences |
Secondary antibodies: | Â | Â | Â | Â |
Anti-rabbit IgG | Alexa Fluor594-conjugated (1:200) | Â | Donkey | A21207, Invitrogen |
Anti-rabbit IgG | Alexa Fluor546-conjugated (1:200) | Â | Goat | A11010, Invitrogen |
Anti-rabbit IgG | Alexa Fluor488-conjugated (1:200) | Â | Goat | A11070, Invitrogen |
Anti-rabbit IgG | Alexa Fluor488-conjugated (1:200) | Â | Chicken | A21441, Invitrogen |
Anti-rat IgG | Alexa Fluor568-conjugated (1:200) | Â | Goat | A11077, Invitrogen |
Anti-rat IgG | Alexa Fluor594-conjugated (1:200) | Â | Goat | A11007, Invitrogen |
Anti-rat Ig | Alexa Fluor488-conjugated (1:200) | Â | Goat | A11006, Invitrogen |
Streptavidin | HRP-conjugated Streptavidin (1:200) | Â | Â | P0397, DakoCytomation |