Double fluorescence staining of FITC-albumin prelabeled rat forebrain tissue sections
| First primary antibodies | First visualizing immunoreagents | Second primary antibodies | Second visualizing immunoreagents |
|---|---|---|---|
| Biotinylated mouse anti-NeuN (1:100; Merck Millipore) | Cy3-streptavidin | Rabbit anti-laminin (1:200; Dakocytomation) | Cy5 donkey anti-rabbit IgG |
| Rabbit anti-laminin (1:400; Dakocytomation) | Cy3-donkey anti-rabbit IgG | Guinea pig anti-GFAP (1:200; Synaptic Systems) | Cy5 donkey anti-guinea pig IgG |
| Biotinylated STL (10 µg/ml; Vector) | Cy3-streptavidin | Guinea pig anti-GFAP (1:200; Synaptic Systems) | Cy5 donkey anti-guinea pig IgG |
| Rabbit anti-collagen IV (1:100; Merck Millipore) | Cy3-donkey anti-rabbit IgG | Biotinylated STL (20 µg/ml; Vector) | Cy5-streptavidin |
| Rabbit anti-AQP4 (1:100; Alomone) | Cy3-donkey anti-rabbit IgG | Biotinylated STL (20 µg/ml; Vector) | Cy5-streptavidin |
| Rabbit anti-AQP4 (1:100; Alomone) | Cy3-donkey anti-rabbit IgG | Guinea pig anti-GFAP (1:200; Synaptic Systems) | Cy5 donkey anti-guinea pig IgG |
| Rabbit anti-S100β (1:600; Synaptic Systems) | Cy3-donkey anti-rabbit IgG | Guinea pig anti-GFAP (1:200; Synaptic Systems) | Cy5 donkey anti-guinea pig IgG |
| Rabbit anti-Iba (1:400; Synaptic Systems) | Cy3-donkey anti-rabbit IgG | Guinea pig anti-GFAP (1:200; Synaptic Systems | Cy5 donkey anti-guinea pig IgG |
| Rabbit anti-Iba (1:200; Synaptic Systems) | Cy3-donkey anti-rabbit IgG | Biotinylated STL (20 µg/ml; Vector) | Cy5-streptavidin |
| Biotinylated mouse anti-CD45c (1:20; Serotec) | Cy3-streptavidin | Rabbit anti-Iba (1:200; Synaptic Systems) | Cy5 donkey anti-rabbit IgG |
| Biotinylated mouse anti-CD8b (1:25; Serotec) | Cy3-streptavidin | Rabbit anti-Iba (1:200; Synaptic Systems) | Cy5 donkey anti-rabbit IgG |
All fluorescent immunoreagents were obtained from Dianova and used at 20 µg/ml for 1 h. AQP4, aquaporin-4; GFAP, glial fibrillary acidic protein; Iba, ionized calcium binding adapter molecule 1; NeuN, neuronal nuclei; STL, Solanum tuberosum agglutinin (potato lectin).