Anti-Aurora A antibody - Centrosome Marker (ab12875)

Overview

  • Product name
    Anti-Aurora A antibody - Centrosome Marker
    See all Aurora A primary antibodies
  • Description
    Rabbit polyclonal to Aurora A - Centrosome Marker
  • Specificity
    ab12875 detects a single clean band of just below 50kD in Jurkat and Hela cell lysates. This band corresponds to the size of Aurora A seen using other antibodies.
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IF, ICCmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human Aurora A.

    (Peptide available as ab13008.)

  • Positive control
    • Jurkat cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab12875 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
IHC-P
WB
ICC/IF
ICC
  • Application notes
    ICC: 1/500.
    IF: 1/500.
    IHC-P: 1/15 - 1/20. Antigen retrieval was performed using either Sodium Citrate or Tris/EDTA before commencing with the IHC staining protocol. Customer feedback suggests that a 1/15 dilution of ab12875 is recommended when using the sodium citrate antigen retreival method and a 1/20 dilution is recommended when using the Tris/EDTA antigen retrieval method.

    WB: 1/500. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).


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

    • Function
      Contributes to the regulation of cell cycle progression. Required for normal mitosis. Associates with the centrosome and the spindle microtubules during mitosis and functions in centrosome maturation, spindle assembly, maintenance of spindle bipolarity, centrosome separation and mitotic checkpoint control. Phosphorylates numerous target proteins, including ARHGEF2, BRCA1, KIF2A, NDEL1, PARD3, PLK1 and BORA. Regulates KIF2A tubulin depolymerase activity (By similarity). Required for normal axon formation. Plays a role in microtubule remodeling during neurite extension. Important for microtubule formation and/or stabilization.
    • Tissue specificity
      Highly expressed in testis and weakly in skeletal muscle, thymus and spleen. Also highly expressed in colon, ovarian, prostate, neuroblastoma, breast and cervical cancer cell lines.
    • Sequence similarities
      Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications
      Activated by phosphorylation at Thr-288; this brings about a change in the conformation of the activation segment. Phosphorylation at Thr-288 varies during the cell cycle and is highest during M phase. Autophosphorylated at Thr-288 upon TPX2 binding. Phosphorylated upon DNA damage, probably by ATM or ATR.
      Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by the anaphase-promoting complex (APC), leading to its degradation by the proteasome.
    • Cellular localization
      Cytoplasm > cytoskeleton > centrosome. Cytoplasm > cytoskeleton > spindle pole. Detected at the neurite hillock in developing neurons (By similarity). Localizes on centrosomes in interphase cells and at each spindle pole in mitosis.
    • Information by UniProt
    • Database links
    • Alternative names
      • AIK antibody
      • ARK-1 antibody
      • ARK1 antibody
      • AURA antibody
      • Aurka antibody
      • Aurora 2 antibody
      • Aurora A antibody
      • Aurora kinase A antibody
      • Aurora-related kinase 1 antibody
      • Aurora/IPL1 like kinase antibody
      • Aurora/IPL1-related kinase 1 antibody
      • AURORA2 antibody
      • Breast tumor-amplified kinase antibody
      • BTAK antibody
      • hARK1 antibody
      • IAK antibody
      • IPL1 related kinase antibody
      • MGC34538 antibody
      • OTTHUMP00000031340 antibody
      • OTTHUMP00000031341 antibody
      • OTTHUMP00000031342 antibody
      • OTTHUMP00000031343 antibody
      • OTTHUMP00000031344 antibody
      • OTTHUMP00000031345 antibody
      • OTTHUMP00000166071 antibody
      • OTTHUMP00000166072 antibody
      • PPP1R47 antibody
      • Protein phosphatase 1, regulatory subunit 47 antibody
      • Serine/threonine kinase 15 antibody
      • Serine/threonine kinase 6 antibody
      • Serine/threonine-protein kinase 15 antibody
      • Serine/threonine-protein kinase 6 antibody
      • Serine/threonine-protein kinase aurora-A antibody
      • STK15 antibody
      • STK6 antibody
      • STK6_HUMAN antibody
      • STK7 antibody
      see all

    Images

    • All lanes : Anti-Aurora A antibody - Centrosome Marker (ab12875) at 1/500 dilution

      Lane 1 : Jurkat whole cell lysate
      Lane 2 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Alexa Fluor 680 Goat anti Rabbit IgG at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size : 46 kDa
      Observed band size : 48 kDa (why is the actual band size different from the predicted?)
    • Metaphase HeLa cells stained with ab12875 and counterstained with DAPI. The antibody shows the localisation of Aurora A to the centrosome during metaphase. 100x magnification.

      Panel 1: DNA stained with DAPI.
      Panel 2: Cells stained with ab12875 (1/500 dilution).
      Panel 3: DAPI staining is shown in red and ab12875 in green.

    • The image shows staining of formalin fixed paraffin embedded human tonsil sections with ab12875 (1/20 working dilution). Clear nuclear staining can be seen.

    References

    This product has been referenced in:
    • Hammond D  et al. Melanoma-associated mutations in protein phosphatase 6 cause chromosome instability and DNA damage owing to dysregulated Aurora-A. J Cell Sci 126:3429-40 (2013). Read more (PubMed: 23729733) »
    • Zeng K  et al. Protein phosphatase 6 regulates mitotic spindle formation by controlling the T-loop phosphorylation state of Aurora A bound to its activator TPX2. J Cell Biol 191:1315-32 (2010). Read more (PubMed: 21187329) »

    See all 5 Publications for this product

    Customer reviews and Q&As

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    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris gel)
    Sample
    Human Cell lysate - whole cell (U2OS-osteosarcoma)
    Specification
    U2OS-osteosarcoma
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Dr. Carlos le Sage

    Verified customer

    Submitted Jan 21 2014

    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
    20 µg
    Specification
    HeLa
    Gel Running Conditions
    Reduced Denaturing (10% SDS PAGE)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 15°C
    Username

    Ms. AeLee Jeong

    Verified customer

    Submitted Apr 04 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cultured Cells (Head and neck cancer cell)
    Specification
    Head and neck cancer cell
    Fixative
    Acetone
    Permeabilization
    No
    Blocking step
    FBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 27°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 04 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Bladder)
    Specification
    Bladder
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
    Username

    Abcam user community

    Verified customer

    Submitted Jan 23 2007

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry
    Sample
    Human Cell (HeLa and SK-N-SH)
    Specification
    HeLa and SK-N-SH
    Fixative
    Paraformaldehyde
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Oct 12 2005

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

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