Overview

  • Product nameAnti-PLK1 antibody [35-206]See all PLK1 primary antibodies ...
  • Description
    Mouse monoclonal [35-206] to PLK1
  • Tested applicationsFlow Cyt, ICC/IF, IHC-P, IP, WB, ICC more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    His-PLK1 full length purified from Sf9 cells.

  • Epitopeaa330-370.

Properties

Applications

Our Abpromise guarantee covers the use of ab17056 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Use 1µg for 106 cells.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent dilution.
IP Use at an assay dependent dilution.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 67 kDa). This clone is superior to clone 36-298 (ab17057) in Western blotting and IP, but suffers from high background in IF and ICC.
ICC Use at an assay dependent dilution.

Target

  • FunctionSerine/threonine-protein kinase that performs several important functions throughout M phase of the cell cycle, including the regulation of centrosome maturation and spindle assembly, the removal of cohesins from chromosome arms, the inactivation of APC/C inhibitors, and the regulation of mitotic exit and cytokinesis. Required for recovery after DNA damage checkpoint and entry into mitosis. Required for kinetochore localization of BUB1B. Phosphorylates SGOL1. Required for spindle pole localization of isoform 3 of SGOL1 and plays a role in regulating its centriole cohesion function. Phosphorylates BORA, and thereby promotes the degradation of BORA. Contributes to the regulation of AURKA function. Regulates TP53 stability through phosphorylation of TOPORS.
  • Tissue specificityPlacenta and colon.
  • Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.
    Contains 2 POLO box domains.
    Contains 1 protein kinase domain.
  • Developmental stageAccumulates to a maximum during the G2 and M phases, declines to a nearly undetectable level following mitosis and throughout G1 phase, and then begins to accumulate again during S phase.
  • Post-translational
    modifications
    Catalytic activity is enhanced by phosphorylation of Thr-210. Phosphorylation at Thr-210 is first detected on centrosomes in the G2 phase of the cell cycle, peaks in prometaphase and gradually disappears from centrosomes during anaphase.
    Autophosphorylation and phosphorylation of Ser-137 may not be significant for the activation of PLK1 during mitosis, but may enhance catalytic activity during recovery after DNA damage checkpoint.
    Ubiquitinated by the anaphase promoting complex/cyclosome (APC/C) in anaphase and following DNA damage, leading to its degradation by the proteasome. Ubiquitination is mediated via its interaction with FZR1/CDH1. Ubiquitination and subsequent degradation prevents entry into mitosis and is essential to maintain an efficient G2 DNA damage checkpoint.
  • Cellular localizationNucleus. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > centrosome. During early stages of mitosis, the phosphorylated form is detected on centrosomes and kinetochores. Localizes to the outer kinetochore. Presence of SGOL1 and interaction with the phosphorylated form of BUB1 is required for the kinetochore localization.
  • Target information above from: UniProt accession P53350 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Cell cycle regulated protein kinase antibody
    • PLK 1 antibody
    • PLK antibody
    • PLK-1 antibody
    • plk1 antibody
    • PLK1_HUMAN antibody
    • polio like kinase antibody
    • Polo like kinase 1 antibody
    • Polo-like kinase 1 antibody
    • Serine/threonine protein kinase 13 antibody
    • Serine/threonine protein kinase PLK1 antibody
    • Serine/threonine-protein kinase 13 antibody
    • Serine/threonine-protein kinase antibody
    • Serine/threonine-protein kinase PLK1 antibody
    • STPK 13 antibody
    • STPK13 antibody
    see all

Anti-PLK1 antibody [35-206] images

  • All lanes : Anti-PLK1 antibody [35-206] (ab17056)

    Lane 1 : Recombinant PLK1
    Lane 2 : U2OS lysate
    Lane 3 : HeLaS3


    Performed under reducing conditions.

    Predicted band size : 67 kDa
    Observed band size : 66 kDa (why is the actual band size different from the predicted?)
  • Immunofluoresence using ab17056 and either U2OS, HeLaS3 or NIH 3T3 cells.

    Plk1 can be seen localising with MKLP in the U2OS cell and with Cdc20 in the HeLaS3 and NIH 3T3 cells shown.

  • Anti-PLK1 antibody [35-206] (ab17056) at 1 µg/ml + A431 cell lysate at 20 µg

    Secondary
    Rabbit-anti mouse Alexa fluor(680) at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 67 kDa
    Observed band size : 66 kDa (why is the actual band size different from the predicted?)
  • ab17056 at 1/100 staining human kidney tubular epithelial cells by ICC/IF. The cells were formaldehyde fixed, permeabilized with Triton X-100 and blocked with goat serum before incuabtion with the antibody. A goat anti-mouse FITC antibody was used as the secondary.

    See Abreview

  • Overlay histogram showing HCT116 cells stained with ab17056 (red line). The cells were fixed with 4% paraformaldehyde (10 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 followed by the antibody (ab17056, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HCT116 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-PLK1 antibody [35-206] (ab17056)

This product has been referenced in:
  • Ragland RL  et al. RNF4 and PLK1 are required for replication fork collapse in ATR-deficient cells. Genes Dev 27:2259-73 (2013). Read more (PubMed: 24142876) »
  • Zhou Z  et al. p53 Suppresses E2F1-dependent PLK1 expression upon DNA damage by forming p53-E2F1-DNA complex. Exp Cell Res N/A:N/A (2013). Read more (PubMed: 24076372) »

See all 8 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-12% MOPS)
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 1µg/mL · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 29 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (Colon carcinoma)
Specification Colon carcinoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Dec 04 2012

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon)
Specification Colon
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer
Permeabilization Yes - Buffer from Dako with Tween20
Username

Mr. Rudolf Jung

Verified customer

Submitted May 06 2011

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (kidney tubular epithelium)
Specification kidney tubular epithelium
Fixative Formaldehyde
Permeabilization Yes - Triton X-100
Blocking step Goat serum (10% in PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Melanie Adler

Verified customer

Submitted Aug 03 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"