Mitochondrial localization of ATP5A using antibody ab119688. Cultured HeLa cells were fixed, permeabilized and then labeled with 15H4C4-FITC (1 µg/ml). Since the antibody is labeled with FITC no secondary antibody is necessary.
Overlay histogram showing HeLa cells stained with ab119688 (blue line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody ab119688 (15H4C4 FITC) at 1µg/1xE6/mL cells for 30 min at 22ºC. Negative control (red line) is unstained d cells. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.