Rats were perfused with cold PBS, cold 4% PFA, and brains were left in 4% PFA overnight. Brains were then cryoprotected with 30% sucrose solution and stored in -80C.
Free floating method was used with 40 micron thick coronal sections. For washes and incubations, 24 well-plate was set on a rotating platform at a speed sufficient enough to keep sections moving in the well.
Secondary antibody was incubated for 2 hrs @ RT with the same diluent used for the primary antibody.
Submitted Dec 30 2014
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