Overview

  • Product nameAnti-PLK1 antibody [36-298]See all PLK1 primary antibodies ...
  • Description
    Mouse monoclonal [36-298] to PLK1
  • Tested applicationsIndirect ELISA, IP, WB, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    His-PLK1 full length purified from Sf9 cells.

  • Epitopeaa330-370.
  • Positive control
    • WB: 293, HeLaS3 or U2OS cell lysate ICC: HeLaS3, NIH 3T3 or U2OS cells
  • General notes

    This antibody is available as an azide-free product (see ab178666). 

Properties

Applications

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 Abreviews Notes
Indirect ELISA 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: 68 kDa).
ICC/IF 1/200. PubMed: 19033445
Flow Cyt Use 1µg for 106 cells.

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 [36-298] images

  • Anti-PLK1 antibody [36-298] (ab17057) at 1 µg/ml + 293 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
  • Immunofluoresence using ab17057 and either HeLaS3, NIH 3T3 or U2OS cells.

  • 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?)
  • 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.
  • Overlay histogram showing HCT116 cells stained with ab17057 (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 (ab17057, 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 IgG1 [ICIGG1] (ab91353, 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 [36-298] (ab17057)

This product has been referenced in:
  • Telkoparan P  et al. Coiled-coil domain containing protein 124 is a novel centrosome and midbody protein that interacts with the Ras-guanine nucleotide exchange factor 1B and is involved in cytokinesis. PLoS One 8:e69289 (2013). ICC/IF ; Human . Read more (PubMed: 23894443) »
  • Tan EP  et al. O-linked N-acetylglucosamine cycling regulates mitotic spindle organization. J Biol Chem 288:27085-99 (2013). Read more (PubMed: 23946484) »

See all 7 Publications for this product

Product Wall

Thank you for contacting us and sorry for the delay in getting back to you.

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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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (rectal cancer)
Specification rectal cancer
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
Permeabilization No
Blocking step Sequential pre-diluted peroxidase (10 min) and protein (10 min) blocks. as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Jul 15 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - PBS TritonX100 (0.5%;10min)
Username

Dr. Kirk McManus

Verified customer

Submitted Jun 28 2007

Thank you for your enquiry. Following discussion with our lab I have determined that the reactivity of this antibody against the phospho- or non-phosphorylated forms of Plk1 has unfortunately not been determined. Should you decide to go ahead an...

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Thank you for your enquiry. According to the EXPASY database PLK1 has been conformed to be expressed in placenta and colon tissue in humans and newborn and adult spleen, fetal and newborn kidney, liver, brain, thymus and adult bone marrow, thymus,...

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Thank you for this information, it is very helpful. Thanks again!

I just wanted to let you know that I was able to obtain some additional information regarding the IF testing with ab17057. For PLK1 staining they usually fixed the cells with PTEMF Buffer according to Kapoor et al, JCB150(5) 2000, 975-988. He mention...

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Thank you for your enquiry. Regarding the IF/ICC images that we have on the online datasheet, at this time we don't have additional details regarding how the cells were fixed. I suspect that fixation with 4% formaldehyde or cold methanol would be ok. I...

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