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Phospho-PLK1 (T210) Recombinant Rabbit Monoclonal Antibody [JJ20-58]

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Specification

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Catalog# ET1701-33

Phospho-PLK1 (T210) Recombinant Rabbit Monoclonal Antibody [JJ20-58]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • FC

  • IHC-P

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

Phospho-PLK1 (T210) Recombinant Rabbit Monoclonal Antibody [JJ20-58]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Thr210 of Human PLK1 aa 181-230 / 603.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IF-Tissue, FC, IHC-P

Molecular Weight

Predicted band size: 68 kDa

Positive Control

HeLa treated with 100nM Nocodazole for 24 hours cell lysate, HeLa treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate, HT-29 treated with 100nM Nocodazole for 24 hours cell lysate, HT-29 treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate, mouse thymus tissue, MCF-7.

Conjugation

unconjugated

Clone Number

JJ20-58

RRID

AB_3070201

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:10-1:50

  • IF-Tissue

  • 1:10-1:50

  • FC

  • 1:10-1:50

  • IHC-P

  • 1:200

Target

Function

Serine/threonine-protein kinase PLK1, also known as polo-like kinase 1 (PLK-1) or serine/threonine-protein kinase 13 (STPK13), is an enzyme that in humans is encoded by the PLK1 (polo-like kinase 1) gene. During interphase, PLK1 localizes to centrosomes. In early mitosis, it associates with mitotic spindle poles. A recombinant GFP-PLK1 protein localizes to centromere/kinetochore region, suggesting a possible role for chromosome separation. Plk1 is an early trigger for G2/M transition. Plk1 supports the functional maturation of the centrosome in late G2/early prophase and establishment of the bipolar spindle. Plk1 phosphorylates and activates cdc25C, a phosphatase that dephosphorylates and activates the cyclinB/cdc2 complex. Plk phosphorylates and activates components of the anaphase-promoting complex (APC).

Background References

1. Platani M et al. Mio depletion links mTOR regulation to Aurora A and Plk1 activation at mitotic centrosomes. J Cell Biol 210:45-62 (2015).

2. Kachaner D et al. Interdomain allosteric regulation of Polo kinase by Aurora B and Map205 is required for cytokinesis. J Cell Biol 207:201-11 (2014).

Sequence Similarity

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.

Tissue Specificity

Placenta and colon.

Post-translational Modification

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. Dephosphorylation at Thr-210 at centrosomes is probably mediated by protein phosphatase 1C (PP1C), via interaction with PPP1R12A/MYPT1. 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. Phosphorylated in vitro by STK10.; 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. Monoubiquitination at Lys-492 by the BCR(KLHL22) ubiquitin ligase complex does not lead to degradation: it promotes PLK1 dissociation from phosphoreceptor proteins and subsequent removal from kinetochores, allowing silencing of the spindle assembly checkpoint (SAC) and chromosome segregation.

Subcellular Location

Nucleus, Chromosome, Cytoplasm, Midbody.

UNIPROT

SwissProt: P53350 Human

SwissProt: Q07832 Mouse

Synonyms

Cell cycle regulated protein kinase antibody

PLK 1 antibody

PLK antibody

PLK-1 antibody

plk1 antibody

PLK1_HUMAN antibody

Polo like kinase 1 antibody

Polo-like kinase 1 antibody

Serine/threonine protein kinase 13 antibody

Serine/threonine protein kinase PLK1 antibody

Expand

Images

  • Western blot analysis of Phospho-PLK1 (T210) on different lysates with Rabbit anti-Phospho-PLK1 (T210) antibody (<a href="/products/ET1701-33" style="font-weight: bold;text-decoration: underline;">ET1701-33</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100nM Nocodazole for 24 hours cell lysate<br />Lane 3: HeLa treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate<br />Lane 4: HT-29 cell lysate<br />Lane 5: HT-29 treated with 100nM Nocodazole for 24 hours cell lysate<br />Lane 6: HT-29 treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 68 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1701-33" style="font-weight: bold;text-decoration: underline;">ET1701-33</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-PLK1 (T210) on different lysates with Rabbit anti-Phospho-PLK1 (T210) antibody (ET1701-33) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100nM Nocodazole for 24 hours cell lysate
    Lane 3: HeLa treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate
    Lane 4: HT-29 cell lysate
    Lane 5: HT-29 treated with 100nM Nocodazole for 24 hours cell lysate
    Lane 6: HT-29 treated with 2mM thymidine for 16 hours then treated with 100nM Nocodazole for 24 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa

    Exposure time: 3 minutes; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-33) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-Phospho-PLK1 (T210) antibody (<a href="/products/ET1701-33" style="font-weight: bold;text-decoration: underline;">ET1701-33</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1701-33" style="font-weight: bold;text-decoration: underline;">ET1701-33</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-Phospho-PLK1 (T210) antibody (ET1701-33) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-33) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Phospho-PLK1 (T210) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1701-33" style="font-weight: bold;text-decoration: underline;">ET1701-33</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Phospho-PLK1 (T210) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-33, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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