Immunocytochemistry of primary neurons

Wash cells with PBS1X/HBSS

Fix cells with paraformaldehyde (PFA) 4% for 0h 30m 0s at room temperature

Wash cells with PBS1X for 0h 5m 0s for three times at room temperature

Permeabilize cells with 0.3% Triton X-100 (in PBS1X) for 0h 5m 0s at room temperature

Wash cells with PBS1X for 0h 5m 0s for three times at room temperature

Saturate cells in blocking solution for at least 0h 30m 0s

Wash cells with PBS1X for 0h 5m 0s for three times at room temperature

Incubate with primary antibody in working solution for 1h 30m 0s at room temperature

Wash cells in working solution for 0h 5m 0s for three times at room temperature

Incubate with secondary antibody in working solution for 1h 0m 0s at room temperature in the dark

Wash cells in working solution for 0h 5m 0s for three times at room temperature

Wash cells in PBS1X for 0h 1m 0s for three times at room temperature

Rinse coverslips in dH₂O

Mount the coverslips on slides with mounting medium