Human E2F3 knockout HeLa cell lysate

• WB

Lab

Western blot - Human E2F3 knockout HeLa cell lysate (AB257415)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : E2F3 knockout HeLa cell lysate 20 μg
Lane 3 : SH-SY5Y cell lysate 20 μg
Lane 4 : U-87 MG cell lysate 20 μg
False colour image of Western blot : Anti-E2F3 antibody staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab152126 was shown to bind specifically to E2F3. A band was observed at 49 kDa in wild-type HeLa cell lysates with no signal observed at this size in E2F3 knockout cell line ab265362 (knockout cell lysate ab257415). To generate this image, wild-type and E2F3 knockout HeLa 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-E2F3 antibody (<a href='/en-us/products/primary-antibodies/e2f3-antibody-ab152126'>ab152126</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

E2F3 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human E2F3 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-e2f3-knockout-hela-cell-line-ab265362'>ab265362</a>)

Lane 3:

SH-SY5Y cell lysate at 20 µg

Lane 4:

U-87 MG cell lysate at 20 µg

Predicted band size: 49 kDa

Observed band size: 49 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human E2F3 knockout HeLa cell lysate (AB257415)

Allele-2 : 19 bp deletion in exon2

• Sanger seq

Unknown

Sanger Sequencing - Human E2F3 knockout HeLa cell lysate (AB257415)

Allele-1 : 38 bp deletion in exon2

• WB

Supplier Data

Western blot - Human E2F3 knockout HeLa cell lysate (AB257415)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The molecular weight observed is consistent with what has been described in the literature (PMID : 14716298, PMID : 36276637). In Western blot, ab320731 was shown to bind specifically to E2F3. Target of interest was observed at 52, 55 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in E2F3 knockout cell line (lane 2) (lane 2, knockout cell line ab265362 / knockout cell lysate ab257415). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-E2F3 antibody [EPR28928-50] (<a href='/en-us/products/primary-antibodies/e2f3-antibody-epr28928-50-ab320731'>ab320731</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human E2F3 knockout HeLa cell lysate (ab257415) at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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: 60 kDa,55 kDa,36 kDa

false

Exposure time: 114s