Anti-Aurora A antibody [35C1]

4 product images

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  Flow Cytometry (Intracellular) - Anti-Aurora A antibody [35C1] (ab13824)

  Overlay histogram showing HeLa cells stained with ab13824 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13824, 2µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (Mouse IgG2b [PLPV219] - Isotype Control ab91366, 2µg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed.

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  Western blot - Anti-Aurora A antibody [35C1] (ab13824)

  This blot was produced using 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200v for 50 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab13824 over night at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

  All lanes: Western blot - Anti-Aurora A antibody [35C1] (ab13824) at 5 µg/mL

  Lane 1: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 20 µg

  Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

  Secondary

  All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 45 kDa

  Observed band size: 125 kDa, 50 kDa, 55 kDa

  Exposure time: 20min

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  Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [35C1] (ab13824)

  This data was developed using the same antibody clone in a different buffer formulation without PBS and sodium azide (ab264552)

  ab264552 staining Aurora A in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab264552 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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  This image is courtesy of a customer review submitted by Dr Kirk McManus

  Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [35C1] (ab13824)

  ab13824, at 1/2000 dilution, detecting Aurora A (green) in Hela Cells in conjunction with a Goat anti-mouse secondary antibody conjugated to Cy3^®. Cells were fixed with methanol and counterstained with DAPI. Please refer to abreview for further details.