Introduction
Methods
Animals
Antibodies
Method | Primary antibody | Secondary antibody |
---|---|---|
Immunogold | AQP4, host: rabbit,Sigma A5971, 1,0 mg/ml diluted 1:400 α-syntrophin (SYN259), host: rabbit, 6 mg/ml diluted 1:200, Gift from Dr. Marvin E. Adams | Goat anti-rabbit 15 nm, Abcam 1:20 Goat anti-rabbit 15 nm, Abcam 1:20 |
Immunofluorescence | AQP4, host: rabbit,Sigma A5971, 1,0 mg/ml diluted 1:400 | Alexa fluor 488, donkey anti-rabbit, Invitrogen, A21206 |
Immunoperoxidase | AQP4, host: rabbit, Sigma A5971; 1,0 mg/ml diluted 1:400 | Biotin Donkey anti-rabbit IgG 31821, Pierce, Thermo Scientific, 1:100 Streptavidin-biotinylated HRP, RPN1051 V, GE Healthcare, 1:100 |
Western blot | AQP4, anti-rabbit,Sigma A5971; 1,0 mg/ml diluted 1:5000 Actin, anti-rabbit, Sigma, A2066; 0,67 mg/ml diluted 1:200 | Mouse monoclonal anti-Rabbit IgG-alkaline phosphatase, A2556, 1:5000 |
Transcardial perfusion fixation
Post-embedding immunogold electron microscopy
Method | Analysed regions and subregions |
---|---|
Immunogold histochemistry | Cortex; parietofrontal cortex, piriform cortex |
Hippocampus; stratum pyramidale, stratum radiatum, dentate gyrus | |
Inferior colliculus | |
Cerebellum; molecular layer, granular layer, white matter | |
Spinal cord (thoracic); white matter, grey matter | |
Quantitative RT-PCR | Parietofrontal cortex, hippocampus, inferior colliculus, cerebellum, whole brain samples |
Western blot | Parietofrontal cortex, inferior colliculus |