Anti-E2F1 antibody [EPR26698-49]

• WB

Lab

Western blot - Anti-E2F1 antibody [EPR26698-49] (AB314311)

Western blot : Anti-E2F1 antibody [EPR26698-49] (ab314311) 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, ab314311 was shown to bind specifically to E2F1. A band was observed at 47 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in E2F1 knockout cell line. To generate this image, wild-type and E2F1 knockout HCT 116 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-E2F1 antibody [EPR26698-49] (ab314311) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

E2F1 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type HEK-293 ab259780 cell lysate at 20 µg

Lane 4:

E2F1 knockout HEK-293 <a href='/en-us/products/cell-lines/human-e2f1-knockout-hek-293-cell-line-ab269502'>ab269502</a> 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

false

• WB

Supplier Data

Western blot - Anti-E2F1 antibody [EPR26698-49] (AB314311)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-E2F1 antibody [EPR26698-49] (ab314311) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human colon cancer tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 70 kDa

true

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-E2F1 antibody [EPR26698-49] (AB314311)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the higher MW band at approximately 140 kDa is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-E2F1 antibody [EPR26698-49] (ab314311) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa transfected with siRNA specifically targeti E2F1 whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney 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: 70 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-E2F1 antibody [EPR26698-49] (AB314311)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab314311 was shown to bind specifically to E2F1. Target of interest was observed at 70 kDa in wild-type HEK-293 cell lysates (lane 1) with no signal observed at this size in E2F1 knockout cell line (lane 2) (lane 2, knockout cell line ab269502 / knockout cell lysate ab281354). The identity of the higher MW band at approximately 140 kDa is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-E2F1 antibody [EPR26698-49] (ab314311) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

E2F1 knockout HEK-293 whole cell lysate at 20 µg

Lane 3:

Raji (human burkitts lymphoma b lymphocyte) 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: 70 kDa

false

Exposure time: 180s