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    aurora-b-antibody-ep1009y-ab45145.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Aurora
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RecombinantRabMAb

Recombinant Anti-Aurora B antibody [EP1009Y] (ab45145)

  • Datasheet
Reviews (1) Submit a question References (20)

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Western blot - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • Immunocytochemistry - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • Flow Cytometry (Intracellular) - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • Immunoprecipitation - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • OI-RD Scanning - Anti-Aurora B antibody [EP1009Y] (ab45145)
  • Anti-Aurora B antibody [EP1009Y] (ab45145)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1009Y] to Aurora B
  • Suitable for: Flow Cyt (Intra), ICC, WB, IHC-P, IP
  • Reacts with: Human

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Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-Aurora B antibody [EP1009Y]
    See all Aurora B primary antibodies
  • Description

    Rabbit monoclonal [EP1009Y] to Aurora B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC, WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Aurora B aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • IP: HeLa cell lysate WB: HeLa cell lysate IHC: human endometrium carcinoma ICC: HeLa cells
  • General notes

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Dissociation constant (KD)

    KD = 5.50 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1009Y
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Aurora
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Phosphorylation
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Aurora
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Aurora

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Aurora B antibody [EP1009Y] (ab197614)
    • Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (ab239837)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human Aurora B protein (ab51435)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab45145 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
Flow Cyt (Intra)
1/300.
ICC
1/20.

For unpurified version use 1/100 - 1/150 dilution

WB (1)
Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.

For unpurified version use at 1/50000 dilution

IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified version use at 1/250-500

IP
Use at an assay dependent concentration.

For unpurified version use at 1/20 dilution

Notes
Flow Cyt (Intra)
1/300.
ICC
1/20.

For unpurified version use 1/100 - 1/150 dilution

WB
Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.

For unpurified version use at 1/50000 dilution

IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified version use at 1/250-500

IP
Use at an assay dependent concentration.

For unpurified version use at 1/20 dilution

Target

  • Function

    May 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 specificity

    High 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 disease

    Note=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 similarities

    Belongs 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 localization

    Nucleus. 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.
  • Target information above from: UniProt accession Q96GD4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9212 Human
    • Omim: 604970 Human
    • SwissProt: Q96GD4 Human
    • Unigene: 442658 Human
    • 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
      • PPP1R48 antibody
      • Protein phosphatase 1 regulatory subunit 48 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

    Images

    • Western blot - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Western blot - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Lanes 1-2 : Anti-Aurora B antibody [EP1009Y] (ab45145) at 1/1000 dilution
      Lanes 3-4 : Anti-Aurora B antibody [EP1009Y] (ab45145) at 1/5000 dilution
      Lanes 5-6 : Anti-Aurora B antibody [EP1009Y] (ab45145) at 1/10000 dilution
      Lanes 7-8 : Anti-Aurora B antibody [EP1009Y] (ab45145) at 1/50000 dilution
      Lanes 9-10 : Anti-Aurora B antibody [EP1009Y] (ab45145) at 1/75000 dilution

      Lanes 1 & 3 & 5 & 7 & 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lanes 2 & 4 & 6 & 8 & 10 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Nocodozole Stimulated

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG VHH Single Domain (HRP) (ab191866) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

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


      Exposure time: 8 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab45145 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

    • Immunocytochemistry - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Immunocytochemistry - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Aurora B with Purified ab45145 at 1:50 dilution (5.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium carcinoma tissue sections labeling Aurora B with purified ab45145 at 1/200 dilution (1.35 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Flow Cytometry (Intracellular) - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Flow Cytometry (Intracellular) - Anti-Aurora B antibody [EP1009Y] (ab45145)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Aurora B with Purified ab45145 at 1/300 dilution (0.1 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left). Unlabeled control - /.

       

    • Immunoprecipitation - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Immunoprecipitation - Anti-Aurora B antibody [EP1009Y] (ab45145)

      Purified ab45145 at 1/20 dilution (1µg) immunoprecipitating Aurora B in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab45145 + HeLa whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45145 in HeLa whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 39 kDa

    • OI-RD Scanning - Anti-Aurora B antibody [EP1009Y] (ab45145)
      OI-RD Scanning - Anti-Aurora B antibody [EP1009Y] (ab45145)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Aurora B antibody [EP1009Y] (ab45145)
      Anti-Aurora B antibody [EP1009Y] (ab45145)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (20)

    Publishing research using ab45145? Please let us know so that we can cite the reference in this datasheet.

    ab45145 has been referenced in 20 publications.

    • Tsunematsu T  et al. APC/CCdh1 is required for the termination of chromosomal passenger complex activity upon mitotic exit. J Cell Sci 133:N/A (2020). PubMed: 32934012
    • Kato S  et al. Cyanidioschyzon merolae aurora kinase phosphorylates evolutionarily conserved sites on its target to regulate mitochondrial division. Commun Biol 2:477 (2019). PubMed: 31886415
    • Xie CM  et al. Cardiac glycoside bufalin blocks cancer cell growth by inhibition of Aurora A and Aurora B activation via PI3K-Akt pathway. Oncotarget 9:13783-13795 (2018). PubMed: 29568394
    • Lin J  et al. FBW7 is associated with prognosis, inhibits malignancies and enhances temozolomide sensitivity in glioblastoma cells. Cancer Sci 109:1001-1011 (2018). WB ; Human . PubMed: 29427543
    • Li R  et al. Serine/Threonine Kinase Unc-51-like Kinase-1 (Ulk1) Phosphorylates the Co-chaperone Cell Division Cycle Protein 37 (Cdc37) and Thereby Disrupts the Stability of Cdc37 Client Proteins. J Biol Chem 292:2830-2841 (2017). PubMed: 28073914
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

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    Western blot abreview for Anti-Aurora B antibody [EP1009Y]

    Good
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Carlos le Sage

    Verified customer

    Submitted Jan 21 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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