Anti-Aurora B (phospho T232) antibody (ab61074)

Overview

  • Product nameAnti-Aurora B (phospho T232) antibody
    See all Aurora B primary antibodies
  • Description
    Rabbit polyclonal to Aurora B (phospho T232)
  • Specificityab61074 detects endogenous levels of AurB only when phosphorylated at threonine 232.
  • Tested applicationsWB, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic phosphopeptide derived from human Aurora B around the phosphorylation site of threonine 232 (R-K-TP-M-C).

  • Positive control
    • Extracts from COS7 cells, starved in serum-free media for 18-24 hours then treated with Nocodazole (1ug/ml in fresg serum-free media) for 16 hours. This antibody gave a positive result in IHC in the following FFPE tissue: Human Testis Seminoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab61074 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
WB 1/500 - 1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
ELISA 1/20000.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • FunctionMay be directly involved in regulating the cleavage of polar spindle microtubules and is a key regulator for the onset of cytokinesis during mitosis. Component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. Phosphorylates 'Ser-10' and 'Ser-28' of histone H3 during mitosis. Required for kinetochore localization of BUB1 and SGOL1. Interacts with INCENP.
  • Tissue specificityHigh level expression seen in the thymus. It is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Expressed during S and G2/M phase and expression is up-regulated in cancer cells during M phase.
  • Involvement in diseaseNote=Disruptive regulation of expression is a possibile mechanism of the perturbation of chromosomal integrity in cancer cells through its dominant-negative effect on cytokinesis.
  • Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Ubiquitinated by different BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complexes. Ubiquitinated by the BCR(KLHL9-KLHL13) E3 ubiquitin ligase complex, ubiquitination leads to removal from mitotic chromosomes and is required for cytokinesis. During anaphase, the BCR(KLHL21) E3 ubiquitin ligase complex recruits the CPC complex from chromosomes to the spindle midzone and mediates the ubiquitination of AURKB. Ubiquitination of AURKB by BCR(KLHL21) E3 ubiquitin ligase complex may not lead to its degradation by the proteasome.
  • Cellular localizationNucleus. Chromosome. Chromosome > centromere. Cytoplasm > cytoskeleton > spindle. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalized with gamma tubulin in the mid-body.
  • Information by UniProt
  • Database links
  • Alternative names
    • AIK2 antibody
    • AIM-1 antibody
    • AIM1 antibody
    • ARK-2 antibody
    • ARK2 antibody
    • AurB antibody
    • AURKB antibody
    • AURKB_HUMAN antibody
    • Aurora 1 antibody
    • Aurora and Ipl1 like midbody associated protein 1 antibody
    • Aurora kinase B antibody
    • Aurora related kinase 2 antibody
    • Aurora- and Ipl1-like midbody-associated protein 1 antibody
    • Aurora-B antibody
    • Aurora-related kinase 2 antibody
    • Aurora/IPL1 related kinase 2 antibody
    • Aurora/IPL1-related kinase 2 antibody
    • IPL1 antibody
    • Serine/theronine kinase 12 antibody
    • Serine/threonine protein kinase 12 antibody
    • Serine/threonine-protein kinase 12 antibody
    • Serine/threonine-protein kinase aurora-B antibody
    • STK-1 antibody
    • STK1 antibody
    • STK12 antibody
    • STK5 antibody
    see all

Anti-Aurora B (phospho T232) antibody images

  • ab61074 staining Aurora B in Mouse 14.5dpc tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/200 in 1% BSA + 1% FBS in TBS) for 16 hours. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • IHC image of Aurora B (phospho T232) staining in Human Testis Seminoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab61074, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Aurora B (phospho T232) antibody (ab61074) at 1/500 dilution

    Lane 1 : Extracts from COS7 cells treated with Nocodazole (1ug/ml, 16hours)
    Lane 2 : Extracts from COS7 cells, treated with Nocodazole (1ug/ml, 16hours) and with the immunising phospho-peptide.


    Predicted band size : 39 kDa
    Observed band size : 39 kDa

References for Anti-Aurora B (phospho T232) antibody (ab61074)

This product has been referenced in:
  • Lin BW  et al. Overexpression of Aurora-C interferes with the spindle checkpoint by promoting the degradation of Aurora-B. Cell Death Dis 5:e1106 (2014). WB ; Human . Read more (PubMed: 24603334) »
  • 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 3 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Mouse Cell (NMuMG)
Specification NMuMG
Permeabilization Yes - PBS Tw-20 1%
Fixative Formaldehyde
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Submitted Nov 27 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Mouse Tissue sections (Mouse 14.5dpc)
Specification Mouse 14.5dpc
Permeabilization No
Fixative Formaldehyde
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Submitted Nov 14 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (15% acrylamide)
Sample Mouse Cell lysate - whole cell (NMuMG)
Specification NMuMG
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Oct 24 2013

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