This data was developed using the same antibody clone in a different buffer formulation (ab238667)

Overlay histogram showing HUVEC cells stained with ab238667 (red line). The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab238667) (1 µg/1x10⁶ cells) for 30 min on ice. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min on ice.

Isotype control antibody (black line) was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Overlay histogram showing HUVEC (Human umbilical vein endothelial cell line) cells stained with ab190147 (red line). The cells were fixed with 4% formaldehyde (10 minutes) 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 (ab190147, 1 µg/1x10⁶ cells) for 30 minutes at 22ºC. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/4000 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1 µg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab190147).

ab190147 stained Malme-3M cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab190147 at 5 µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor^® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab190147).