Anti-Ajuba antibody

• IP

Supplier Data

Immunoprecipitation - Anti-Ajuba antibody (AB241957)

Ajuba was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab241957 at 6 μg/reaction. Western blot was performed from the immunoprecipitate using ab241957 at 1 μg/ml.

Lane 1 : ab241957 IP in HeLa whole cell lysate.
Lane 2 : Control IgG IP in HeLa whole cell lysate.

Detection : Chemiluminescence with exposure time of 3 minutes.

Lysates prepared in NETN buffer.

All lanes:

Immunoprecipitation - Anti-Ajuba antibody (ab241957)

Predicted band size: 57 kDa

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• WB

Supplier Data

Western blot - Anti-Ajuba antibody (AB241957)

Lysates prepared in NETN buffer.

All lanes:

Western blot - Anti-Ajuba antibody (ab241957) at 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

Predicted band size: 57 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-Ajuba antibody (AB241957)

Western blot : Anti-AJUBA antibody (ab241957) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab241957 was shown to bind specifically to AJUBA. A band was observed at 55-60 kDa in wild-type A549 cell lysates with no signal observed at this size in AJUBA knockout cell line. To generate this image, wild-type and AJUBA 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Ajuba antibody (ab241957) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

AJUBA knockout A549 cell lysate at 20 µg

Lane 3:

HCT 116 cell lysate at 20 µg

Lane 4:

MEF1 cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 57 kDa

Observed band size: 55 kDa,60 kDa

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