Anti-PLK1 antibody [Mixed clones, 3G251] (ab14210)

Overview

  • Product nameAnti-PLK1 antibody [Mixed clones, 3G251]
    See all PLK1 primary antibodies
  • Description
    Mouse monoclonal [Mixed clones, 3G251] to PLK1
  • SpecificityThis antibody reacts specifically with the 68 kDa Plk1 protein. However, cross-reactivity with a 110 kDa protein of unknown identity is sometimes observed.
  • Tested applicationsICC/IF, Functional Studies, ELISA, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis
    Predicted to work with: Pig
  • Immunogen

    Fusion protein:

    DPACIPIFWVS KWVDYSDKYG LGYQLCDNS VGVLFNDST RLILYNDGDSL QYIERDGTESY LTVSSHPNSLM KKITLLKYFRN YMSEHLLKAGA NITPREGDELA RLPYLRTWF RTRSAIILH LSNGSVQINFF QDHTKLILCP LMAAVTYIDE KRDFRTYRLS LLEEYGCCKE LASRLRYARTM VDKLLSSRSAS NRLKAS

    , corresponding to amino acids 402-603 of Human Plk1.

  • Positive control
    • HeLa, A431 and NIH 3T3 cells.
  • General notesIsotype is IgG1 kappa and IgG2b kappa. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.


    ab14210 has been shown to preferentially detect centrosomal-associated Plk1. An alternative antibody, Rabbit polyclonal to Plk1 (ab14209) can be used to detect kinetochore-associated Plk1.

Properties

Applications

Our Abpromise guarantee covers the use of ab14210 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
Functional Studies Use at an assay dependent dilution. PubMed: 20427271In vitro kinase assay.
ELISA Use a concentration of 0.1 - 1 µg/ml.
IP Use a concentration of 2 - 5 µg/ml.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 67 kDa).

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.
  • 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 antibody
    • Serine/threonine-protein kinase 13 antibody
    • Serine/threonine-protein kinase PLK1 antibody
    • STPK 13 antibody
    • STPK13 antibody
    see all

Anti-PLK1 antibody [Mixed clones, 3G251] images

  • Human U2OS cells were fixed for 10 minutes in 3.7% formaldehyde (30mM sucrose) and permeabilized for 5 minutes in PBS-Triton (0.1%). The fixed cells were blocked for 30 minutes then incubated for 40 minutes with 3µg/ml anti-Pkl1, ab14210, (green) and anti-Centrin (red). The DNA was counterstained with DAPI (blue). All incubations were carried out at room temperature.

    The staining pattern shows that ab14210 localizes to the centrosomes during most of mitosis (upper panel) and the midbody during telophase (lower panel). The centrosomal pool of Plk1 can be seen by comparing with centrin, a centriolar marker. An alternative antibody, Rabbit polyclonal to Plk1 (ab14209) can be used to selectively stain the kinetochore-associated Plk1.


  • Human U2OS cells were fixed and permeabilized in 4% formaldehyde, 0.2% Triton X100.The fixed cells were blocked for 30 minutes then incubated for 40 minutes with 3µg/ml anti-Pkl1, ab14210, (green) and anti-Centrin (red). The DNA was counterstained with DAPI (blue). All incubations were carried out at romm temperature.

    The staining pattern shows that ab14210 localizes to the centrosomes during most of mitosis (upper panel) and the midbody during telophase (lower panel). The centrosomal pool of Plk1 can be seen by comparing with centrin, a centriolar marker. An alternative antibody, Rabbit polyclonal to Plk1 (ab14209) can be used to selectively stain kinetochore-associated Plk1.

References for Anti-PLK1 antibody [Mixed clones, 3G251] (ab14210)

This product has been referenced in:
  • Castiel A  et al. The Stil protein regulates centrosome integrity and mitosis through suppression of Chfr. J Cell Sci 124:532-9 (2011). WB ; Mouse . Read more (PubMed: 21245198) »
  • Ciciarello M  et al. Nuclear reformation after mitosis requires downregulation of the Ran GTPase effector RanBP1 in mammalian cells. Chromosoma 119:651-68 (2010). WB ; Human . Read more (PubMed: 20658144) »

See all 6 Publications for this product

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As requested, I have looked into the sequence homology between the immunogen used to raise antibodies mentioned and the relevant canineprotein sequence. I have to men...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 300000 cells
Specification HeLa
Treatment RNAi specific to plk1 72h
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Jun 05 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (colo205 and HT29)
Specification colo205 and HT29
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Jul 29 2005

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