Rabbit anti cytokeratin 5 alexa fluor 647

Dissociated cells from mouse prostate were stained with rat anti-CD49f-PE (BioLegend), rat anti-CD326 (EpCAM)-APC/Cy7 (BioLegend) and goat anti-Trop2-APC (R & D Systems) for 20 min on ice. Cells were washed with PBS and fixed in 1ml of 2% paraformaldehyde made from 16% paraformaldehyde (Electron Microscopy Sciences) in PBS for 15 min on ice. Cells were washed with PBS and permeabilized in 1ml of permeabilization buffer (0.1% Saponin (Sigma-Aldrich), 5% FBS (Corning) in PBS) for 15 min at room temperature in the dark. Cells were resuspended in 100 μL of permeabilization buffer and stained with either rabbit anti-cytokeratin 5-Alexa Fluor 647 (Abcam) and rabbit anti-cytokeratin 8-Alexa Fluor 488 (Abcam), mouse anti-cytokeratin 14-FITC (Abcam), mouse anti-cytokeratin 18-FITC (Abcam) or rat anti-Ki67-FITC (BioLegend) for 20 min at room temperature in the dark. Cells were washed and resuspended in permeabilization buffer for analysis on a BD FACS Canto (BD Biosciences).