All lanes : Anti-Beta Arrestin 2 antibody (ab54790) at 1 µg/ml

Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : ARRB2 knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Human Lung tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Observed band size: 50 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab54790 observed at 50 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

ab54790 was shown to react with Beta Arrestin 2 in wild-type HepG2 cells in Western blot with loss of signal observed in ARRB2 knockout cell line ab262320 (knockout cell lysate ab257283). Wild-type HepG2 and ARRB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab54790 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Beta Arrestin 2 antibody (ab54790) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human tonsil.

This image was generated using the ascites version of the product.

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

Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : ARRB2 knockout HepG2 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : K562 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Observed band size: 55 kDa why is the actual band size different from the predicted?

Lanes 1- 4: Merged signal (red and green). Green - ab54790 observed at 55 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

 ab54790 was shown to react with ARRB2 in wild-type HepG2 cells in western blot. Loss of signal was observed when knockout cell line ab262320 (knockout cell lysate ab257283) was used. Wild-type HepG2 and ARRB2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab54790 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

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.

Observed band size: 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.

 

All lanes : Anti-Beta Arrestin 2 antibody (ab54790)

Lane 1 : Beta Arrestin 2 transfected 293T
Lane 2 : Non- transfected 293T

Western blot against tagged recombinant protein immunogen using ab54790 Beta Arrestin 2 antibody at 1ug/ml. Predicted band size of immunogen is 35 kDa

This image was generated using the ascites version of the product.

ab54790 at 10 ug/ml staining Beta Arrestin 2 in human Hela cells by Immunocytochemistry / Immunofluorescence.

This image was generated using the ascites version of the product.

Overlay histogram showing HeLa cells stained with ab54790 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 (ab54790, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG2a [ICIGG2A], (ab91361, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.