All lanes : Anti-NAPE-PLD antibody [OTI3D5] (ab119259) at 1/2000 dilution

Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate transfected with pCMV6-ENTRY control cDNA
Lane 2 : HEK-293T cell lysate transfected with pCMV6-ENTRY NAPE-PLD cDNA

Lysates/proteins at 5 µg per lane.

Predicted band size: 46 kDa

Overlay histogram showing SHSY-5Y (Human neuroblastoma cell line from bone marrow) cells stained with ab119259 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab119259, 1/1000 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x10⁶ cells) used under the same conditions. Unlabeled 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 SHSY-5Y cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.

Paraffin-embedded human endometrium adenocarcinoma tissue stained for NAPE-PLD with ab119259 at a 1/150 dilution in immunohistochemical analysis. Heat-induced epitope retrieval by 10 mM citric buffer, pH 6.0, 100°C for 10 minutes.

Paraffin-embedded human bladder carcinoma tissue stained for NAPE-PLD with ab119259 at a 1/150 dilution in immunohistochemical analysis. Heat-induced epitope retrieval by 10 mM citric buffer, pH 6.0, 100°C for 10 minutes.

Paraffin-embedded human tonsil tissue stained for NAPE-PLD with ab119259 at a 1/150 dilution in immunohistochemical analysis. Heat-induced epitope retrieval by 10 mM citric buffer, pH 6.0, 100°C for 10 minutes.