Recombinant
RabMAb

Recombinant Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)

Rabbit recombinant monoclonal Aurora A antibody [EPR5026]. Validated in WB, Flow Cyt, ICC/IF and tested in Mouse, Human. Independently reviewed in 1 review(s).

Reviews (1)Q&A (1)

Overview

  • Product name

    Anti-Aurora A antibody [EPR5026] - Centrosome Marker
    See all Aurora A primary antibodies

  • Description

    Rabbit monoclonal [EPR5026] to Aurora A - Centrosome Marker

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IPmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Aurora A. The exact sequence is proprietary.

  • Positive control

    • WB: BXPC-3, LnCaP, SKBR-3 and HepG2 cell lysates. ICC/IF: HeLa cells.

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab108353 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 1/1000 - 1/10000. Predicted molecular weight: 46 kDa.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/100 - 1/250.
IP 1/60.
  • Application notes
    Is unsuitable for IHC-P.

    • 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-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 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

      • Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        All lanes : Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353) at 1/1000 dilution

        Lane 1 : BXPC-3 cell lysate
        Lane 2 : LnCaP cell lysate
        Lane 3 : SKBR-3 cell lysate
        Lane 4 : HepG2 cell lysate

        Lysates/proteins at 10 µg per lane.

        Predicted band size: 46 kDa

      • Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Immunofluorescent staining of HeLa cells using ab108353 at 1/100
      • Immunoprecipitation - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Immunoprecipitation - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)

        Lane 1: Neuro2a (Mouse neuroblastoma neuroblast) whole cell lysate, 10µg
        Lane 2:         Neuro2a cell lysate 350µg and ab109518 2µg
        Lane 3:         Neuro2a cell lysate, 350µg and rabbit IgG (ab172730) , 2µg

        Purified ab108353 immunoprecipitating Aurora A in Neuro2a cell lysates. Primary antibody was used at a 1/60 dilution (20 µg/ml).  For western blotting, ab108353 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

        Blocking and diluting buffer used: 5% NFDM/TBST.

      • Immunoprecipitation - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Immunoprecipitation - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)

        Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
        Lane 2:         HepG2 cell lysate 350µg and ab109518 2µg
        Lane 3:          HepG2 cell lysate 350µg and rabbit IgG (ab172730) , 2µg

        Purified ab108353 immunoprecipitating Aurora A in Neuro2a cell lysates. Primary antibody was used at a 1/60 dilution (20 µg/ml).  For western blotting, ab108353 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

        Blocking and diluting buffer used: 5% NFDM/TBST.

      • Flow Cytometry - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Flow Cytometry - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Overlay histogram showing HeLa cells stained with ab108353 (red line). The cells were fixed with 80% methanol (5 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 (ab108353, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
      • Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353) at 1/1000 dilution + Neuro-2a cell lysate at 10 µg

        Predicted band size: 46 kDa

      • Other - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Other - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD

      References

      ab108353 has not yet been referenced specifically in any publications.

      Publishing research using ab108353? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      1-2 of 2 Abreviews or Q&A

      Western blot abreview for Anti-Aurora A [EPR5026] antibody - Centrosome Marker

       Inconclusive
      Abreviews
      abreview image
      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
      Read More

      Dr. Carlos le Sage

      Verified customer

      Submitted Jan 21 2014

      Question

      My technician forwarded your note below to me and I simply do not understand your response. You say that the antibody noted below is the best bet as the peptide has a fair amount of alignment between human and Chlamy Aurora A. Well, although the two proteins have 67% homology, there is no overlap at all that I can detect between the human peptide immunogen and any part of the Chlamy Aurora A sequence.

      Have you made a mistake, or posted an incorrect ab link? I am at a loss for understanding your suggested ab (and if I hadn't looked, I would be at a loss for about $*** as well).

      thanks,

      Read More
      Answer

      Thank you for your reply.

      I apologize for the confusion, there was a difference in the numerical order of amino acids listed in the alignment I was performing. You are correct, ab115883 should not be chosen as the immunogen sequence does not overlap with the Chlamy Aurora A sequence.

      I reran the search using the proper numerical organization and came up with the following antibody:
      ab61114 (https://www.abcam.com/Aurora-A-antibody-ab61114.html): the immunogen sequence is around the human Aurora A amino acid 288. The human and Chlamy sequences are quite homologous in this region, as amino acids 285 and 288 are the only two mismatched amino acids from 275-301.

      I'm actually confident enough in the homology within this region to offer our Abtrial program for testing ab61114 in your Chlamy sequences. Information regarding this program is given below:

      To our knowledge, ab61114 has not been tested in C. reinhardtii. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

      Simply follow these easy steps below to apply for our AbTrial Program:

      1. Reply to this email, letting us know you are interested in testing this product.

      2. Our scientists will email you an inactive personal discount code for the value of the product.

      3. Purchase and test the product at the regular price.

      4. Submit your results, including your discount code in the additional notes section of your Abreview.

      5. Once the Abreview is submitted, the discount code will become active.

      6. Apply your discount code on your next order to receive that value off.

      The Terms and Conditions of this offer can be found at: https://www.abcam.com/abtrial

      Additionally, until 12/16/12 we are running a rabbit monoclonal antibody promotion, our "RabMab" promotion, which entitles you to a free rabbit monoclonal antibody with purchase of a primary antibody. I found a rabbit monoclonal antibody ab108353 (https://www.abcam.com/Aurora-A-antibody-EPR5026-ab108353.html) that has 54% homology with the Chlamy sequence and might be worth a try, otherwise you could choose a different Rabbit monoclonal antibody that you desire. Details of the RabMab promotion are given below:
      https://www.abcam.com/index.html?pageconfig=resource&rid=15447

      Please let me know if you have any questions about this offer and I would be happy to help you further.

      Read More

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