All lanes : Anti-Beta Arrestin 2 antibody (ab54790) at 1/500 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ARRB2 knockout HEK293T cell lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : Human lung tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 46 kDa
Observed band size: 48-55 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab54790 observed at 48-55 kDa. Red - loading control, ab181602 observed at 37 kDa.

ab54790 Anti-Beta Arrestin 2 antibody was shown to specifically react with Beta Arrestin 2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266116 (knockout cell lysate ab257282) was used. Wild-type and Beta Arrestin 2 knockout samples were subjected to SDS-PAGE. ab54790 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 1 bp deletion in exon4

Representative images of ARRB2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.