Mouse anti human ve cadherin antibody

HCAECs plated on glass coverslips were cultured to ~90% confluence before being transfected with hP2Y₂R-eGFP cDNA. The cell transfectants were maintained in growth medium for 72 h and transferred to serum-free medium for 12 h. Then, cells were incubated with or without 100 µM UTP for 5 min at 37°C, washed in ice-cold PBS, fixed for 10 min in 4% (w/v) paraformaldehyde, treated with 0.1% (v/v) Triton X-100 for 5 min, and rinsed in PBS. Fixed cells were incubated with mouse anti-human VE-cadherin antibody (1:100 dilution, BD Bioscience, CA) for 1 h, washed and stained with Alexa Fluor 594 goat anti-mouse IgG (1:200 dilution, Invitrogen) for 1 h. Coverslips were mounted on glass slides in ProLong antifade reagent (Life Technologies, Grand Island, NY) and examined using a Zeiss inverted LSM 510 META confocal laser scanning microscope (CLSM) equipped with a C Apochromat 40× objective. Images were acquired, processed and analyzed with a Zeiss LSM Image Examiner.