Anti-BRCA2 antibody [EPR23442-43]

• WB

Lab

Western blot - Anti-BRCA2 antibody [EPR23442-43] (AB239375)

Western blot : Anti-SHP1 antibody [Y476] ab32559 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 68 kDa in Wild-type MCF7 cell lysates with a band observed at 55 kDa in PTPN6 CRISPR-Cas9 Edited MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-BRCA2 antibody [EPR23442-43] (ab239375) at 1/500 dilution

Lane 1:

Wild-type MCF7 at 20 µg

Lane 2:

BRCA2 knockout MCF7

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 270 kDa,400 kDa

Observed band size: 270 kDa,400 kDa

false

• WB

Lab

Western blot - Anti-BRCA2 antibody [EPR23442-43] (AB239375)

Negative control : Capan-1 (PMID : 16251802). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-BRCA2 antibody [EPR23442-43] (ab239375)

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 100 µg

Lane 2:

Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 100 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 1/10000 dilution

Predicted band size: 384 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-BRCA2 antibody [EPR23442-43] (AB239375)

Anti-BRCA2 antibody [EPR23442-43] (ab239375) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab239375 was shown to bind specifically to BRCA2. A band was observed at 290 kDa in wild-type MCF7 cell lysates with no signal observed at this size in BRCA2 knockout cell line ab286285 (knockout cell lysate ab300210). To generate this image, wild-type and BRCA2 knockout MCF7 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-BRCA2 antibody [EPR23442-43] (ab239375) at 1/500 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

Western blot - Human BRCA2 knockout MCF7 cell lysate (ab300210) at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

Lane 5:

HT-29 cell lysate at 20 µg

Secondary

Lanes 1 - 5:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 5:

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

Observed band size: 290 kDa

false