Anti-BCOR antibody

3 product images

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  Western blot - Anti-BCOR antibody (ab245423)

  All lanes: Western blot - Anti-BCOR antibody (ab245423) at 0.04 µg/mL

  Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

  Lane 2: HeLa whole cell lysate at 15 µg

  Lane 3: HeLa whole cell lysate at 5 µg

  Developed using the ECL technique.

  Predicted band size: 192 kDa

  Exposure time: 30s

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  Immunoprecipitation - Anti-BCOR antibody (ab245423)

  BCOR was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab245423 at 3 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab245423 at 1 μg/ml.
  

  Lane 1: ab245423 IP in HeLa whole cell lysate.

  Lane 2: Control IgG IP in HeLa whole cell lysate.

  Detection: Chemiluminescence with an exposure time of 3 seconds.

  All lanes: Immunoprecipitation - Anti-BCOR antibody (ab245423)

  Predicted band size: 192 kDa

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  Western blot - Anti-BCOR antibody (ab245423)

  False colour image of Western blot: Anti-BCOR antibody staining at 1/1000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab245423 was shown to bind specifically to BCOR. A band was observed at 192 kDa in wild-type HeLa cell lysates with no signal observed at this size in BCOR CRISPR-Cas9 edited cell line Human BCOR knockout HeLa cell line ab265125 (CRISPR-Cas9 edited cell lysate Human BCOR knockout HeLa cell lysate ab258327). The band observed in the CRISPR-Cas9 edited lysate lane below 192 kDa is likely to represent a truncated form of BCOR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and BCOR CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA 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 before development with Optiblot (ECL reagent Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-BCOR antibody (ab245423) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: BCOR CRISPR-Cas9 edited HeLa cell lysate at 20 µg

  Lane 3: K562 cell lysate at 20 µg

  Lane 4: Caco-2 cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 192 kDa