Goat anti rat alexafluor 568

OCT-embedded tissues were sectioned at 12 μm or 20 μm. Sections were rehydrated and blocked in antibody wash (5% heat inactivated goat serum, 0.1% Triton X-100 in Tris-Buffered Saline) for 45 minutes. Tissues were incubated with primary antibodies overnight at 4°C, washed three times with 0.1% Triton X-100 in Tris-Buffered Saline (TBST), and incubated with secondary antibodies for 1 hour at room temperature. Tissues were washed three times with TBST and incubated with Hoechst 33342 for 5 minutes. Slides were coverslipped with ProLong Gold (ThermoFisher) mounting media and imaged on a BioTek Lionheart FX microscope. Primary antibodies used were: mouse anti-ARL13B (1:1000, NeuroMab, N295B/66); rabbit anti-acetylated α-tubulin (1:1000,Cell Signaling, 5335); mouse anti-acetylated α-tubulin (1:2500, Sigma, T7451); chicken anti-ACIII (1:1000, Encor, CPCA-ACIII); rat anti-insulin (1:1000, R&D systems, MAB1417); rabbit anti-glucagon (1:2000, Abcam, ab92517). Secondary antibodies used were goat anti-mouse AlexaFluor 488, goat anti-chicken AlexaFluor 647, goat anti-rat AlexaFluor 568 and donkey anti-rabbit AlexaFluor 555 (1:500, ThermoFisher).