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

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Overview

Product name

Anti-PLK1 antibody [36-298]
See all PLK1 products (21) ...

Description

Mouse monoclonal [36-298] to PLK1

Tested applications

Indirect ELISA, IP, WB, ICC/IFmore details

Cross reactivity

Reacts with

Mouse, Rat, Human

Immunogen

His-PLK1 full length purified from Sf9 cells.

Epitope

aa330-370.

Positive control

WB: 293, HeLaS3 or U2OS cell lysate ICC: HeLaS3, NIH 3T3 or U2OS cells

Properties

Form

Liquid

Storage instructions

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.05% Sodium Azide
Constituents: PBS

Concentration

Concentration information loading...

Purity

IgG fraction

Clonality

Monoclonal

Clone number

36-298

Isotype

IgG1

Research areas

Cancer >> Cell cycle >> Cell division
Epigenetics and Nuclear Signaling >> Cell cycle >> Kinases/Phosphatases >> Other
Cell Biology >> Cell Cycle >> Cell Division >> Spindle
Cell Biology >> Cell Cycle >> Kinases/Phosphatases >> Other

  • Western blot - Plk1 antibody [36-298] (ab17057)Western blot - Plk1 antibody [36-298] (ab17057) image (enlarge)

  • Immunofluorescence - Plk1 antibody [36-298] (ab17057)Immunofluorescence - Plk1 antibody [36-298] (ab17057) image (enlarge)

  • Western blot - Plk1 antibody [36-298] (ab17057)Western blot - Plk1 antibody [36-298] (ab17057) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab17057 in the following tested applications.

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

Application notes

ICC/IF: 1/200 (PMID 19033445).
IP: Use at an assay dependent dilution.
I-ELISA: Use at an assay dependent dilution.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 68 kDa).


Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Target

Function

Serine/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 specificity

Placenta and colon.

Sequence similarities

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.
Contains 2 POLO box domains.
Contains 1 protein kinase domain.

Developmental stage

Accumulates 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 localization

Nucleus. 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 accessionP53350 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

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

Database links

see all

Anti-PLK1 antibody [36-298] images:

  Western blot - Plk1 antibody [36-298] (ab17057)

Western blot - Plk1 antibody [36-298] (ab17057)

Anti-PLK1 antibody [36-298] (ab17057) at 1 µg/ml + 293 cell lysate at 20 µg

Secondary
Rabbit polyclonal Secondary Antibody to 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

  Immunofluorescence - Plk1 antibody [36-298] (ab17057)

Immunofluorescence - Plk1 antibody [36-298] (ab17057)

Immunofluoresence using ab17057 and either HeLaS3, NIH 3T3 or U2OS cells.

  Western blot - Plk1 antibody [36-298] (ab17057)

Western blot - Plk1 antibody [36-298] (ab17057)

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

Lane 1 : recombinant Plk1
Lane 2 : U2OS cell extract
Lane 3 : HeLaS3 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?)

  Immunofluorescence - Plk1 antibody [36-298] (ab17057)

Immunofluorescence - Plk1 antibody [36-298] (ab17057)

In panel one HeLa 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 min. with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC 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.

Scott Slattery and Mike Mancini

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

This product has been referenced in:

  • Ward A  et al. Aberrant methylation of Polo-like kinase CpG islands in Plk4 heterozygous mice. BMC Cancer 11:71 (2011). WB; Mouse.Read more (PubMed: 21324136) »
  • Shang ZF  et al. Inactivation of DNA-dependent protein kinase leads to spindle disruption and mitotic catastrophe with attenuated checkpoint protein 2 Phosphorylation in response to DNA damage. Cancer Res 70:3657-66 (2010). WB; Human.Read more (PubMed: 20406977) »

See all 5 publications for this product

Publishing research using ab17057? Please let us know so that we can cite the reference in this datasheet

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