Anti-TAZ antibody

3 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAZ antibody (ab110239)

  IHC image of TAZ staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110239, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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  Western blot - Anti-TAZ antibody (ab110239)

  TAZ Western blot staining using rabbit Anti-TAZ antibody

  False colour image of Western blot: Anti-TAZ antibody staining at 1 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab110239 was shown to bind specifically to TAZ. A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in WWTR1 knockout cell line Human WWTR1 knockout HeLa cell line ab281598 (knockout cell lysate ab282950). To generate this image, wild-type and WWTR1 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 5 % 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.}

  All lanes: Western blot - Anti-TAZ antibody (ab110239) at 1 µg/mL

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

  Lane 2: WWTR1 knockout HeLa cell lysate at 20 µg

  Lane 3: A549 cell lysate at 20 µg

  Lane 4: K562 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

  Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 44 kDa

  Observed band size: 52 kDa, 37 kDa

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  Western blot - Anti-TAZ antibody (ab110239)

  All lanes: Western blot - Anti-TAZ antibody (ab110239) at 1 µg/mL

  Lane 1: A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 20 µg

  Lane 2: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

  Lane 3: A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 20 µg with Immunising peptide

  Lane 4: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg with Immunising peptide

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 44 kDa

  Observed band size: 36 kDa, 41 kDa, 53 kDa

  Exposure time: 4min