Anti-BCL9L antibody [EPR27491-35]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BCL9L antibody [EPR27491-35] (AB314440)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SW480 (human colorectal adenocarcinoma epithelial cell) cells labelling BCL9L with ab314440 at 1/100 (5.13 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1011 (2ug/ml) dilution (Green). Confocal image showing nuclear staining in SW480 cell line. Negative control : RKO (PMID : 24419084). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1011 (2ug/ml) dilution.

• WB

Lab

Western blot - Anti-BCL9L antibody [EPR27491-35] (AB314440)

Western blot : Anti-BCL9L antibody [EPR27491-35] (ab314440) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab314440 was shown to bind specifically to BCL9L. A band was observed at 190 kDa in wild-type A549 cell lysates with no signal observed at this size in BCL9L knockout cell line. To generate this image, wild-type and BCL9L 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-BCL9L antibody [EPR27491-35] (ab314440) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

Wild-type A549 cell lysate at 20 µg

Lane 3:

Wild-type A549 cell lysate at 10 µg

Lane 4:

BCL9L knockout A549 cell lysate at 30 µg

Lane 5:

HeLa cell lysate at 20 µg

Lane 6:

HEK-293 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

Observed band size: 190 kDa

false

• WB

Supplier Data

Western blot - Anti-BCL9L antibody [EPR27491-35] (AB314440)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands between 75 kDa and 160 kDa are unknown. In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-BCL9L antibody [EPR27491-35] (ab314440) at 1/1000 dilution

All lanes:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 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 1/20000 dilution

Observed band size: 190 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-BCL9L antibody [EPR27491-35] (AB314440)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : RKO (PMID : 24419084). The identity of the bands between 75 kDa and 160 kDa are unknown. In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-BCL9L antibody [EPR27491-35] (ab314440) at 1/1000 dilution

Lane 1:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

RKO (human colon epithelial cell) whole cell lysate at 20 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 1/20000 dilution

Observed band size: 190 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-BCL9L antibody [EPR27491-35] (AB314440)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the band at 160 kDa is unknown. In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-BCL9L antibody [EPR27491-35] (ab314440) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti BCL9L whole cell lysate at 20 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 1/20000 dilution

Observed band size: 190 kDa

false

Exposure time: 180s