Anti-GAB1 antibody [EPR375]

• IP

Unknown

Immunoprecipitation - Anti-GAB1 antibody [EPR375] (AB133486)

ab133486 (purified) at 1/80 immunoprecipitating GAB1 in HEK293 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/10,000) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-GAB1 antibody [EPR375] (ab133486)

Predicted band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-GAB1 antibody [EPR375] (AB133486)

Blocking buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-GAB1 antibody [EPR375] (ab133486) at 1/2000 dilution

All lanes:

K562 cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-GAB1 antibody [EPR375] (AB133486)

Blocking buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-GAB1 antibody [EPR375] (ab133486) at 1/2000 dilution

All lanes:

HEK293 cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 77 kDa

false

• WB

Lab

Western blot - Anti-GAB1 antibody [EPR375] (AB133486)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : GAB1 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : Hek293 whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133486 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab133486 was shown to recognize GAB1 in wild-type HAP1 cells as signal was lost at the expected MW in GAB1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GAB1 knockout samples were subjected to SDS-PAGE. ab133486 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GAB1 antibody [EPR375] (ab133486)

Predicted band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-GAB1 antibody [EPR375] (AB133486)

All lanes:

Western blot - Anti-GAB1 antibody [EPR375] (ab133486) at 1/1000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

293T cell lysate at 10 µg

Lane 4:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 77 kDa

false