Anti-ATF6 antibody [1-7]

• WB

Lab

Western blot - Anti-ATF6 antibody [1-7] (AB122897)

Lane 1 : Wild type HAP1 whole cell lysate (50 μg)
Lane 2 : ATF6 knockout HAP1 whole cell lysate (50 μg)
Lanes 1 - 2 : Merged signal (red and green). Green - ab122897 observed at 95 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab122897 was shown to specifically recact with ATF6 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when ATF6 knockout samples were used. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. ab122897 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution respectively. Blots were developed 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 1/10,000 dilution for 1hr at room temperature before imaging.

All lanes:

Western blot - Anti-ATF6 antibody [1-7] (ab122897)

Predicted band size: 74 kDa

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• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ATF6 antibody [1-7] (AB122897)

Immunofluorescent analysis of HeLa cells using ab122897 at a dilution of 1/100. The cells were fixed with paraformaldehyde. The secondary antibody was an Alexa Fluor^® 488 conjugated goat anti-mouse IgG at a dilution of 1/1000. The antibody staining is shown in the top panel, and the merged image with DAPI counter-staining is shown in the lower panel.

• WB

Unknown

Western blot - Anti-ATF6 antibody [1-7] (AB122897)

Conversion of precursors pATF6(P) to pATF6(N) in DTT- or tunicamycin-treated cells. ATF6 is constitutively expressed as pATF6a(P) (~90-kDa protein), and converted to pATF6a(N) (>50-kDa protein) in ER-stressed cells.

All lanes:

Western blot - Anti-ATF6 antibody [1-7] (ab122897) at 1/500 dilution

Lane 1:

HeLa cell extract: control

Lane 2:

HeLa cell extract: treated with 1mM DTT for 1 hour

Lane 3:

HeLa cell extract: treated with 2 μg/ml tunicamycin for 3 hour

Lane 4:

HeLa cell extract: treated with 2 μg/ml tunicamycin for 7 hour

Lane 5:

293 cell extract: control

Lane 6:

293 cell extract: treated with 1mM DTT for 1 hour

Lane 7:

293 cell extract: treated with 2 μg/ml tunicamycin for 3 hour

Lane 8:

293 cell extract: treated with 2 μg/ml tunicamycin for 7 hour

Predicted band size: 74 kDa

false

• WB

Unknown

Western blot - Anti-ATF6 antibody [1-7] (AB122897)

Conversion of precursors pATF6(P) to pATF6(N) in DTT- or tunicamycin-treated cells. ATF6 is constitutively expressed as pATF6a(P) (~90-kDa protein), and converted to pATF6a(N) (>50-kDa protein) in ER-stressed cells.

All lanes:

Western blot - Anti-ATF6 antibody [1-7] (ab122897) at 1/500 dilution

Lane 1:

293T cell lysate: control

Lane 2:

293T cell lysate: treated with 2 μg/ml tunicamycin for 7 hours

Lane 3:

293T cell lysate: treated with 1mM DTT for 1 hour.

Lane 4:

HeLa S3 Tet-off cell lysate: control

Lane 5:

HeLa S3 Tet-off cell lysate: treated with 1mM DTT for 1 hour.

Predicted band size: 74 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-ATF6 antibody [1-7] (AB122897)

ATF6 was detected by Western blot (Input; lanes 1, 2, 7, and 8) using ab122897 (No. 1-7). After immunoprecipitation (IP) with non-related IgG (IP; lanes 3, 4, 9, and 10) or ab122897 (No. 1-7) (IP; lanes 5, 6, 11, and 12), samples were subjected to SDS-PAGE and analyzed by Western blot using ab122897 (No. 1-7) and anti-mouse IgG antibody (light chain specific).

All lanes:

Immunoprecipitation - Anti-ATF6 antibody [1-7] (ab122897)

Predicted band size: 74 kDa

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