Anti MAP2, Guinea Pig Protocol

1. Perfusion fixation	Perfuse with sodium citrate/PBS to remove blood, followed by perfusion with 4% PFA.
2. Post-fixation	4% PFA (24 hours)
3. Immersion in sucrose	30% Sucrose/PBS (4℃, overnight ~ approximately 2 days)
4. Preparation of frozen sections	After removing sucrose, prepare frozen blocks and cut 30-μm thick sections using a cryostat.
5. Glycine treatment	25 mM glycine/PBS (approximately 20 minutes)
6. Blocking	5% normal goat serum in 0.3% Triton X-100/PBS (4℃, overnight)
7. Primary antibody reaction	Anti MAP2, Guinea pig (1:1,500), 1% normal goat serum in 0.3% Triton X-100/PBS (4℃, overnight ~ 2 days).
8. Washing	0.3% Triton X-100/PBS (5 minutes x 3)
9. Secondary antibody reaction	Alexa Fluor 488 AffiniPure Goat Anti-Guinea Pig IgG (H+L) (1:500) (Jackson ImmunoResearch, Product Number: 106-545-003), 1% normal goat serum in 0.3% Triton X-100/PBS (room temperature, 2 hours)
10. Washing	0.3% Triton X-100/PBS (5 minutes x 3)
11. Mounting	Store at 4℃ in a dark place.