Anti-FAT/FAT1 antibody

• WB

Supplier Data

Western blot - Anti-FAT/FAT1 antibody (AB241372)

Prepared using NETN lysis buffer.

All lanes:

Western blot - Anti-FAT/FAT1 antibody (ab241372) at 0.1 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 50 µg

Lane 4:

TCMK-1 (Mouse kidney epithelial cell line) whole cell lysate at 50 µg

Lane 5:

NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 50 µg

Predicted band size: 506 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-FAT/FAT1 antibody (AB241372)

Western blot : Anti-FAT1 antibody (ab241372) staining at 1/2000 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab241372 was shown to bind specifically to FAT1. A band was observed at 506 kDa in wild-type A549 cell lysates with no signal observed at this size in FAT1 knockout cell line. To generate this image, wild-type and FAT1 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 3 % milk 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 before development with a high-sensitivity ECL substrate kit and imaged with 3 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-FAT/FAT1 antibody (ab241372) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human FAT1 Knockout A549 cell line (ab314987)

Lane 2:

FAT1 knockout A549 cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HUVEC cell lysate at 20 µg

Secondary

Lanes 1 - 4:

HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 506 kDa

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