Ab68545

Pancreatic tissues were excised from newborn piglets and immediately frozen in OCT with liquid N2. Tissue segments were kept at −80 °C. Tissues were cut into 8 µm sections, fixed in 10% cold Z-fix for 5 min, permeabilized in 0.2% Triton X-100 for 10 min and blocked in 10% goat serum for 30 min at room temperature. Tissue sections were incubated for 1 hour at 37°C with anti-beta-III tubulin (1:250 dilution, Millipore MAB1637, Temecula, CA), anti-chromogranin A (1:400 dilution, Abcam ab43861, Cambridge, MA), anti-von Willebrand (1:1000 dilution, Abcam ab68545, Cambridge, MA), protein gene product 9.5 (1:100 dilution, Abcam ab8189, Cambridge, MA) followed by secondary antibody (Alexa-flour 488 and Alexa-flour 568 (1:500 dilution, Life Technologies, Grand Island, NY) for 30 min at room temperature. Sections were mounted with Vectashield containing DAPI (Vector Labs, Burlingame, CA) to visualize nuclei. Images were acquired using identical parameters on a Zeiss 710 confocal microscope.