Overlay histogram showing HEK293 cells stained with ab140630 (red 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 (ab140630, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.

All lanes : Anti-OXGR1/GPR99 antibody [EPR6305(2)] (ab140630) at 1/1000 dilution

Lane 1 : A549 cell lysate
Lane 2 : NIH 3T3 cell lysate
Lane 3 : Fetal brain tissue lysate
Lane 4 : Fetal kidney tissue lysate
Lane 5 : Placenta tissue lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 38 kDa

Immunofluorescent staining of NIH 3T3 cells labelling OXGR1/GPR99 with ab140630 at 1/250 dilution.