Staining

BrdU labelling - http://www.lbl.gov/LBL-Programs/lifesciences/BissellLab/labprotocols/brdu.htm Bisell Lab. BrdU labelling for human cells Using Boehringer Mahnein Kit cat. #1296 736, or Roche kit cat # 1 296 736. (Same kit) Not provided with the kit: Cell culture media, 50mM glycine, IS blocking solution and DAPI. 1-Grow cells on appropriate substrate and under appropriate condition. 2-Aspirate cell culture medium and add BrdU labelling medium: - [Read more]

Occludine staining - http://www.lbl.gov/LBL-Programs/lifesciences/BissellLab/labprotocols/occludin.htm Bisell Lab. For Occludine staining : Wash 1X with PBS -Fixation : formalin 15 min at RT -Wash 2X with PBS 1X -Triton 0.5%, for 5min -Wash 1X with PBS 1X, 5min -Wash 3X with PBS/glycine 1X, 10min -Blocking with GS 10% in IF (plus Fab#152 for 30 min) -Primary AB : dilute 1/50 in GS 10% (plus Fab#152), at least 2 hrs or O/N (4 degree) (ask johanne for the antibody) - [Read more]