All lanes : Anti-P4HB antibody [EPR9499] (ab137110) at 1/2000 dilutionLane 1 : Mouse brainLane 2 : Mouse heartLane 3 : Mouse kidneyLane 4 : Mouse spleen

Overlay histogram showing HepG2 cells stained with ab137110 (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 (ab137110, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human breast carcinoma tissue, staining P4HB using ab137110 at a 1/100 dilution.

Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human brain tissue, staining P4HB using ab137110 at a 1/100 dilution.

Immunofluorescence analysis of HepG2 cells, staining P4HB (green) using ab137110 at a 1/250 dilution. Cell nuclei are shown in blue.