Rat anti ve cadherin antibody

For immunohistochemistry, brain sections were washed with 0.1 M phosphate buffer (PB) and then incubated for 30 minutes at room temperature in a blocking solution containing TNB, TBST (0.1%) and normal goat serum (3%). For detection of vascular endothelial cells, sections were incubated overnight at 4 °C with monoclonal rat anti-CD31 antibody (BD Biosciences, 1:50). Tight junction proteins were detected with the following antibodies: mouse anti-Claudin-5 antibody (1:200, ThermoFischer); rat anti-VE-Cadherin antibody (1:100, ThermoFischer) and rabbit anti-ZO-1 antibody (1:100, ThermoFischer). Pericytes were visualized with goat anti-CD13 (1:200; R&D). The primary antibody incubation was followed by 2 h incubation at room temperature with corresponding fluorescence secondary antibodies (1:500, ThermoFisher). DAPI (1:2000 in 0.1 M PB, Sigma) was used to visualize nuclei. Sections were mounted in 0.1 M PB on Superfrost PlusTM microscope slides and coverslipped using Mowiol.

Brain sections were washed with 0.1M phosphate buffer (PB) and incubated with blocking solution containing donkey serum (10%) in PB for 30 min at room temperature. For detection of vascular endothelial cells, sections were incubated overnight at 4°C with monoclonal rat anti-CD31 antibody (BD Biosciences, 1:50). The localization of tight/adherens junction proteins was assessed using the following antibodies: mouse anti-Claudin-5 antibody (1:200, Thermo Fisher Scientific); rat anti-VE-Cadherin antibody (1:100, Thermo Fisher Scientific), and rabbit anti-ZO-1 antibody (1:100, Thermo Fisher Scientific). Pericyte coverage was visualized with goat anti-CD13 (1:200; R&D Systems). The primary antibody incubation was followed by 2 h incubation at room temperature with corresponding fluorescent secondary antibodies (1:500, Thermo Fisher Scientific). Nuclei were counterstained with DAPI (1:2,000 in 0.1 M PB, Sigma). Sections were mounted in 0.1 M PB on Superfrost PlusTM microscope slides and coverslipped using Mowiol.