Table 3.
Primary and secondary antibodies used in the histological analyses
| Antibody | Dilution | Incubation time | Source | Marker of | Application |
|---|---|---|---|---|---|
| Primary antibodies | |||||
| Rabbit-anti-tyrosine hydroxylase | 1:1000 | Overnight | Pel-Freeze, Rogers, AR, USA | Tyrosine hydroxylase Dopaminergic neurons | Rat IHC, FF |
| Rat-anti-BrdU | 1:100 | 36h | Serotec, UK | Proliferating cells | Rat IF, FF |
| Rabbit-anti-laminin | 1:100 | 36h | Sigma-Aldrich, Sweden | Endothelial basal lamina | Rat IF, FF |
| Mouse-anti-nestin | 1:5000 | Overnight | BD Pharmingen, | Immature endothelium | Rat IHC, FF |
| Sheep-anti-albumin | 1:60000 1:5000 | Overnight | Biogenesis, USA | Albumin extravasation | Rat IHC, FF Human IHC, PF |
| Mouse-anti-endothelial barrier antigen (EBA) | 1:1000 | Overnight | Covance, USA | Endothelium with intact BBB properties | Rat IHC, FF Rat IF, PF |
| Rabbit-anti-VEGF (A-20) | 1:30 1:100 | Overnight | Santa Cruz, USA | Vascular endothelial growth factor (VEGF) | Rat IF/IHC PF Human IHC, PF |
| Mouse-anti-GFAP | 1:100 | Overnight | Millipore | Glial fibrillary acidic protein, astrocytes | Rat IF, FF/PF |
| Mouse-anti-CD11b (OX-42) | 1:100 | Overnight | Serotec, UK | Microglia | Rat IF, FF |
| Mouse-anti-CD68 (ED-1) | 1:100 | Overnight | Serotec, UK | Activated microglia | Rat IF, FF |
| Mouse-anti-CD34 | 1:300 | Overnight | Serotec, UK | Endothelium | Human IHC, PF |
| Mouse-anti-human nestin | 1:750 | Overnight | Millipore | Immature endothelium | Human IHC, PF |
| Secondary antibodies | |||||
| Biotinylated-goat-anti-rabbit/Biotinylated-horse-anti-mouse | 1:200 | 2 h | Vector Laboratories | Rat IF, FF / Human IHC, PF | |
| Cy3-donkey-anti-rat Cy2/Cy3-donkey-anti-mouse (IgG and IgM specific, respectively) Cy5-donkey-anti-mouse | 1:200 | 2 h | Jackson Immunoresearch | Rat IF, PF, FF |
BBB = blood brain barrier; BrdU = bromodeoxyuridine; FF = free-floating; IF = immunofluorescence; IHC = bright-field immunohistochemistry; PF = paraffin-embedded sections.