Anti β tubulin isotype 3 antibody

Before immunolabeling, dewaxed and rehydrated sections were blocked for 1 h with 1% Bovine Serum Albumin (BSA, cat nr: 9048‐46‐8, Merck) and 0.1% Triton X‐100 (Tx, cat nr: 9002‐93‐1, Sigma‐Aldrich) in phosphate‐buffered saline (PBS) pH 7.4. Then, slices were incubated overnight at 4°C with primary antibodies in PBS, 0.1% BSA, 0.001% Tx: anti‐β‐Tubulin Isotype III antibody (1:500, cat nr: 302302, Synaptic Systems); anti‐β‐Tubulin Isotype III antibody (1:500, cat nr: T5076, Sigma‐Aldrich); anti‐GAD67 (Gad1; 1:200, cat nr: MA5‐31377, Invitrogen); anti‐GluR2 (Gria2; 1:200, cat nr: PA5‐19496, Invitrogen); and anti‐CaMKIV (Camk4; 1:200, cat nr: PA1‐542, Invitrogen), respectively. Subsequently, slices were incubated for 2.5 h at RT with secondary antibodies in PBS, 0.1% BSA (1:2000 AlexaFluor 633 goat anti‐mouse cat nr: A21046 and 1:2000 AlexaFluor 488 goat anti‐rabbit, cat nr: A11034, Invitrogen; 1:2000 AlexaFluor 633 goat anti‐rabbit, cat nr: A21071, Invitrogen and 1:2000 AlexaFluor 488 goat anti‐mouse, cat nr: A11001, Invitrogen), respectively. Finally, glass slides were mounted with Fluoroshield with DAPI (cat nr: F6057, Sigma‐Aldrich). Between each step of the immunostaining procedure, sections were washed with PBS (3 × 10 min, RT).