Table 1.
Antibodies applied for flow cytometry, immunohistochemistry and fluorescence stainings.
| Cell population | Antibody | Isotype | Species of origin | Cat. # |
| Flow cytometry: | ||||
| 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* |
| FITC-conjugated | IgG2b | Rat | 556923, BD Biosciences | |
| PE-conjugated | IgG1 | Hamster | 554711, BD Biosciences | |
| APC-conjugated | IgG2 | Hamster | 558141, BC Biosciences | |
| Un-conjugated | Ig fraction | Rabbit | X0903, DakoCytomation | |
| Secondary antibodies: | ||||
| Anti-rabbit I | Alexa Fluor647-conjugated | IgG | Goat | A21244, Invitrogen |
| Anti-rabbit II | Alexa Fluor488-conjugated | IgG | Chicken | A21441, Invitrogen |
| Immunohistochemistry: | ||||
| BM-derived cells | GFP (1:1000) | Rabbit | ab290, Abcam | |
| TNF producing cells | TNF (1:50) | Rat | MM-350, Endogen | |
| TNF producing cells | TNF (1:200) | 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 | |
| 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) | Rabbit | A0082, DakoCytomation | |
| Endothelial | PECAM-1/CD31 (1:100) | IgG2a | Rat | 557355, BD Biosciences |
| Astrocytes | GFAP (1:1,200) | Rabbit | Z0334, DakoCytomation | |
| Astrocytes | GFAP (1:50) | Goat | sc-6170, Santa Cruz | |
| Biotechnology | ||||
| Isotype/serum controls: | Un-conjugated | IgG2b | Rat | IG-851125, Biosite |
| Un-conjugated | IgG2a | Rat | P54605M, Biosite | |
| Un-conjugated | Serum | Goat | X0907, DakoCytomation | |
| Un-conjugated | Ig fraction | Rabbit | X0903, DakoCytomation | |
| Biotin-conjugated | IgG2a,κ | Mouse | 553455, BD Biosciences | |
| Secondary antibodies: | ||||
| Anti-rabbit IgG | Alexa Fluor594-conjugated (1:200) | Donkey | A21207, Invitrogen | |
| Anti-rabbit IgG | Alexa Fluor546-conjugated (1:200) | Goat | A11010, Invitrogen | |
| Anti-rabbit IgG | Alexa Fluor488-conjugated (1:200) | Goat | A11070, Invitrogen | |
| Anti-rabbit IgG | Alexa Fluor488-conjugated (1:200) | Chicken | A21441, Invitrogen | |
| Anti-rat IgG | Alexa Fluor568-conjugated (1:200) | Goat | A11077, Invitrogen | |
| Anti-rat IgG | Alexa Fluor594-conjugated (1:200) | Goat | A11007, Invitrogen | |
| Anti-rat Ig | Alexa Fluor488-conjugated (1:200) | Goat | A11006, Invitrogen | |
| Streptavidin | HRP-conjugated Streptavidin (1:200) | P0397, DakoCytomation | ||
PE: phycoerythrin, PerCP-Cy5.5: perinidin chlorophyll protein-cyanine5.5, APC: allophycocyanin, FITC: fluorescein isothiocyanate, vWF: von Willebrandt's Factor, PECAM-1: platelet endothelial adhesion molecule-1, GFAP: glial fibrillary acidic protein, GFP: green fluorescent protein.