Great Antibody for Flow IF

3T3 cells were plated on glass coverslips and then directly transfected, or alternatively transfected with a later split onto coverslips. Cells were fixed with 4% paraformaldehyde 15 min, permeabilized with 0.2% Triton X-100 in PBS for 10 min, blocked with 5% goat serum and 0.5% BSA in DPBS for 1 hour. Primary antibodies were added overnight at 4°C, 1:500. The samples were washed three times with DPBS, and secondary antibodies applied for one hour at room temperature. After three additional washes with DPBS, the coverslips were mounted with Mounting Medium with DAPI. This antibody works great for flow immunofluorescence.