Anti-Integrin beta 4 antibody [M126] (ab29042)

Overlay histogram showing A431 cells stained with ab29042 (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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab29042, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 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.

Immunofluorescence analysis of rat 804G cells, staining Integrin beta 4 with ab29042.

Cells were fixed in 4% formaldehyde in PBS for 8 minutes, washed thoroughly in PBS, and permeabilized in 0.1% Triton X-100 in PBS for 10 minutes. Samples were incubated with primary antibody at 37°C for 1 hour. The cells on coverslips were washed with PBS, and AlexaFluor®597-conjugated secondary antibodies were applied for 1 hour at 37°C.