Guinea pig anti arc

Free-floating sections were washed several times in 50 mM Tris-buffered saline (TBS) containing 0.1% Triton X-100, incubated in a blocking buffer (0.5% Triton X-100, 5% bovine serum albumin (BSA) in 50 mM TBS) for 30 min at room temperature (RT) and subsequently incubated with the appropriate primary antibody (diluted in 0.1% Triton X-100, 1% BSA in 0.05 M TBS) for 2 days at RT. Guinea pig anti-Arc (1 mg/mL, 1:1000, Synaptic Systems), guinea pig anti-SP (1 mg/mL, 1:2000, Synaptic Systems), rabbit anti-SP (1 mg/ml, 1:2000, Synaptic Systems), rabbit anti-Calbindin D 28k (1 mg/mL, 1:500, Swant), guinea pig anti-AnkG (100 μL lyophilized, 1:500, Synaptic Systems), rabbit anti-α-Actinin2 (0.142 mg/mL, 1:500, Abcam, clone EP2529Y), and rabbit anti-GRP78 (1 mg/mL, 1:500, Biotrend) were used as primary antibodies. After several washing steps, sections were incubated with the appropriate secondary Alexa-conjugated antibodies (2 mg/mL, 1:2000, goat anti guinea pig or goat anti rabbit Alexa Fluor 488 or 568, Invitrogen) for 4 h at RT, counterstained with Draq5 (1:10 000, Thermo Fisher Scientific) to visualize nuclei and finally mounted in Fluorescence Mounting Medium (Dako, Agilent Technologies).