重组Anti-BRCA2抗体[EPR23442-43]

• 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

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• 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='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 384 kDa

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Exposure time: 180s