Specific anti ve cadherin antibody

Heart tissues were collected after the mice were killed, fixed in 4% paraformaldehyde, embedded in tissue processing medium (O.C.T.), and cut horizontally at a 10-mm thickness. The slides were stained with specific anti–VE-cadherin antibody (1:200 dilution, sc-9,989; Santa Cruz Biotechnology, Santa Cruz, CA) at 4°C for overnight. Human umbilical vein endothelial cells obtained from in vitro cell culture were fixed with 3.7% formaldehyde for 20 min and permeabilized with 0.1% Triton X-100 for 20 min, followed by blocking with 3% bovine serum albumin for 30 min at room temperature. Subsequently, cells were incubated with anti–VE-cadherin antibody (1:200 dilution, sc-9,989; Santa Cruz Biotechnology) at 4°C for overnight. Alexa Fluor 594 secondary antibody (1:500 dilution in PBS, A-11005; Invitrogen) was added to the section to visualize the staining. DAPI (blue) was used to counterstain the nuclei. The stained sections and cells were measured using Confocal Microscope (Leica Camera, Wetzlar, Germany).