Immunofluorescent staining of U87 MG cells labelling NPY5R with ab133757 at 1/250.

Overlay histogram showing SHSY-5Y cells stained with ab133757 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab133757, 1/10000 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) (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 result in 4% paraformaldehyde (10 min) fixed SHSY-5Y cells used under the same conditions. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

All lanes : Anti-NPY5R antibody [EPR8382] (ab133757) at 1/10000 dilutionLane 1 : Human hypothalamus lysateLane 2 : Human fetal brain lysateLane 3 : Ramos cell lysateLane 4 : U87 MG cell lysateLane 5 : Caco 2 cell lysateLane 6 : BxPC 3 cell lysateLysates/proteins at 10 µg per lane.SecondaryGoat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Equilibrium disassociation constant (KD)Learn more about KD Click here to learn more about KD