Vectashield anti fade medium with dapi h 1200

Tissue sections were rehydrated through an ethanol gradient and then treated with 6% H₂O₂ in methanol to block the endogenous peroxidase activity. Antigen retrieval was performed with 10 mM citric acid buffer (pH 6.0) at 95°C for 30 minutes. After samples were cooled down to room temperature (RT), they were blocked at RT for two hours followed by incubation with primary antibodies at 4°C overnight. Then, samples were incubated with secondary antibodies conjugated with Alexa Fluor 488 (green) or Alexa Fluor 594 (red) at RT for two hours. After washing, samples were mounted with Vectashield anti–fade medium with DAPI (H-1200, Vector Laboratories). Images were acquired by a Zeiss LSM510 confocal microscope equipped with LSM 510 Version 4.2 SP1 acquisition software (Carl Zeiss). Antibodies: Cx43 (Santa Cruz Biotechnology, sc-9059); E-Cadherin [41 (link)]; pY489-β-catenin (Developmental Studies Hybridoma Bank).