Anti-Beta Arrestin 2 antibody [3G1]

• Flow Cyt

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Flow Cytometry - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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.

• WB

Lab

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

Western blot : Anti-ARRB2 antibody [3G1] (ab54790) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab54790 was shown to bind specifically to ARRB2. A band was observed at 45 kDa in wild-type A549 cell lysates with no signal observed at this size in ARRB2 knockout cell line. To generate this image, wild-type and ARRB2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790) at 1 µg/mL

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ARRB2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-a549-cell-line-ab277840'>ab277840</a>)

Lane 3:

Wild-type HepG2 <a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-hep-g2-cell-line-ab262320'>ab262320</a> cell lysate at 20 µg

Lane 4:

ARRB2 knockout HepG2 <a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-hep-g2-cell-line-ab262320'>ab262320</a> cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 45 kDa

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• WB

Lab

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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.

All lanes:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790) at 1 µg/mL

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

ARRB2 knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-hep-g2-cell-line-ab262320'>ab262320</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Human Lung tissue lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 50 kDa

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• WB

Lab

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790) at 1/500 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

ARRB2 knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-hep-g2-cell-line-ab262320'>ab262320</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 55 kDa

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• WB

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Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ARRB2 knockout HEK293T cell lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Human lung tissue lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 48-55 kDa

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• WB

Lab

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ARRB2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-arrb2-beta-arrestin-2-knockout-hek-293t-cell-line-ab266116'>ab266116</a>)

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Human lung tissue lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 55 kDa

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• WB

Unknown

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (AB54790)

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.

All lanes:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (ab54790)

Predicted band size: 46 kDa

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