Overlay histogram showing A549 cells stained with ab73021 (red line). The cells were fixed with 80% methanol (5 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 (ab73021, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A549 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

ab73021 staining NAPSIN A in cells from human broncho-alveolar lavage by Immunocytochemistry.Cells were fixed in formaldehyde, permeabilized using Triton, blocked with 5% normal horse serum for 1 hour at room temperature and then incubated with ab73021 at a 1/100 dilution for 16 hours. The secondary used was a biotin conjugated horse anti-mouse polyclonal used at a 1/300 dilution.See Abreview