Anti-Integrin beta 1 抗体 [EPR16896] - BSA and Azide free

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (AB250101)

This data was developed using ab179472, the same antibody clone in a different buffer formulation.

Western blot : Anti-Integrin beta 1 antibody [EPR16896] ab179472 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 100-130 kDa in Wild-type A549 cell lysates with no signal observed at this size in ITGB1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc BSA in TBS-0.1 % Tween^® 20 (TBS-T) 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-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EPR16896] (<a href='/products/primary-antibodies/integrin-beta-1-antibody-epr16896-ab179472'>ab179472</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human ITGB1 knockout A549 cell line (<a href='/products/cell-lines/human-itgb1-knockout-a549-cell-line-ab286593'>ab286593</a>) at 20 µg

Lane 3:

Wild-type HCT 116 at 20 µg

Lane 4:

ITGB1 knockout HCT 116 cell line at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 88 kDa

Observed band size: 100-130 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (AB250101)

This data was developed using the same antibody clone in a different buffer formulation (ab179472).

Lanes 1- 2 : Merged signal (red and green). Green - ab179472 observed at 140-150 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab179472 was shown to react with ITGB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255354 (knockout cell lysate ab263847) was used. Wild-type HeLa and ITGB1 knockout HeLa 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. ab179472 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EPR16896] (<a href='/products/primary-antibodies/integrin-beta-1-antibody-epr16896-ab179472'>ab179472</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ITGB1 knockout HeLa cell lysate at 20 µg

Predicted band size: 130 kDa,88 kDa

Observed band size: 140-150 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (AB250101)

This data was developed using the same antibody clone in a different buffer formulation (ab179472).

Lanes 1 - 2 : Merged signal (red and green). Green - ab179472 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab179472 was shown to react with Integrin beta 1 in HCT 116 wild-type cells in western blot with loss of signal observed in ITGB1 knockout cell line ab273724 (ITGB1 knockout cell lysate ab275251). HCT 116 wild-type and ITGB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab179472 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (ab250101) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

ITGB1 knockout HCT116 cell lysate at 20 µg

Predicted band size: 88 kDa

Observed band size: 130 kDa

false

• WB

Supplier Data

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (AB250101)

This data was developed using the same antibody clone in a different buffer formulation (ab179472).

Lanes 1 - 2 : Merged signal (red and green). Green - ab179472 observed at 125 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab179472 was shown to specifically react with Integrin beta 1 in wild-type HAP1 cells as signal was lost in Integrin beta 1 knockout cells. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab179472 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EPR16896] - BSA and Azide free (ab250101) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

Integrin beta 1 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 88 kDa

false