Protocol for immunostaining M.Aihara

this protocol is for neuron, astroglia and microglia but easily applicable for any samples

following procedures are done in room temperature

previous required materials and equipments (available from MA's stock)

marker                        1st                           2nd wave length
neuron anti-MAP2(mono)        100× anti-mouse IgG-TRITC   50× 510-560nm
astroglia anti-GFAP(mono)     100× anti-mouse IgG-TRITC   50× 510-560nm
microglia isolectin B4-FITC   100× 450-490nm

(These antibodys and anti-rabbit IgG-FITC antibody are available from my stock! Please ask me.)


note: you had better to use Fuji PROVIA iso 1600 film.
If you can see easily fluorescent image, exposure time should be 2 or 3 minutes.
If you can see it with difficulty because of weak fluorescence, exposure time should be over 4 minutes.
Try and error!