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Anti-PLK1 antibody [36-298] (ab17057)

Key features and details

  • Mouse monoclonal [36-298] to PLK1
  • Suitable for: ICC, WB, Flow Cyt
  • Reacts with: Mouse, Human, Recombinant fragment
  • Isotype: IgG1

Overview

  • Product name

    Anti-PLK1 antibody [36-298]
    See all PLK1 primary antibodies

  • Description

    Mouse monoclonal [36-298] to PLK1

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Mouse
    Human
    WB
    Human
    Recombinant fragment
    See all applications and species data

  • Immunogen

    Recombinant full length protein corresponding to Human PLK1. His-PLK1 full length purified from Sf9 cells.
    Database link: P53350

  • Epitope

    aa330-370.

  • Positive control

    • WB: HEK-293, HeLa S3 or U-2 OS cell lysate ICC: HeLa S3, NIH/3T3 or U-2 OS cells Flow Cytometry: U-2 0S cells

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-PLK1 antibody [36-298] (ab17057)
    Western blot - Anti-PLK1 antibody [36-298] (ab17057)
    All lanes : Anti-PLK1 antibody [36-298] (ab17057)

    Lane 1 : Recombinant PLK1
    Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) cell extract
    Lane 3 : HeLa S3 cell extract

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 66 kDa
    why is the actual band size different from the predicted?



    10% SDS-PAGE gel.

  • Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)
    Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)

    Immunofluoresence using ab17057 and either HeLa S3, NIH/3T3 (Mouse embyro fibroblast cell line) or U-2 OS (Human bone osteosarcoma epithelial cell line) cells.

  • Western blot - Anti-PLK1 antibody [36-298] (ab17057)
    Western blot - Anti-PLK1 antibody [36-298] (ab17057)
    Anti-PLK1 antibody [36-298] (ab17057) at 1 µg/ml + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg

    Secondary
    Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 66 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes
  • Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)
    Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057) This image is courtesy of Scott Slattery and Mike Mancini

    In panel one HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were stained with ab17057 (green) and DAPI. In the second panel, cells were stained with ab17057 (green) and SH-CREST (red), which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeabilized for 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4°C diluted 1/400 in 5% milk in TBST. Secondary antibody incubated 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen.

    Notes: Ample washing between each step.

    TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.

  • Flow Cytometry - Anti-PLK1 antibody [36-298] (ab17057)
    Flow Cytometry - Anti-PLK1 antibody [36-298] (ab17057)

    Overlay histogram showing U-2 0S (Human bone osteosarcoma epithelial cell line) cells stained with ab17057 (red line). The cells were fixed with 80% methanol (10 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab17057, 1 µg/1 x 106 cells) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150117) at 1/2000 dilution for 30 minutes at 22°C.

    Isotype control antibody (black line) was mouse IgG1 [15-6E10A7] (ab170190, 1 µg/1 x 106 cells) used under the same conditions.

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

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