Anti-Aurora B 抗体 [mAbcam 3609] (ab3609)

Reviews (4)Q&A (2)References (18)

Key features and details

  • Mouse monoclonal [mAbcam 3609] to Aurora B
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Rat, Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Aurora B antibody [mAbcam 3609]
    Aurora B 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [mAbcam 3609] to Aurora B

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, ICC/IF, IHC-Pmore details

  • 種交差性

    交差種: Rat, Human
    交差が予測される動物種: Mouse

  • 免疫原

    Synthetic peptide corresponding to Human Aurora B aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab13569)

  • ポジティブ･コントロール

    • Hela whole cell lysate

  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3609 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa). Block with 1% BSA for 1 hour.
ICC/IF 1/50.
IHC-P Use at an assay dependent concentration.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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.

  • 関連疾患

    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.

  • 配列類似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
    Contains 1 protein kinase domain.

  • 翻訳後修飾

    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.

  • 細胞内局在

    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.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-Aurora B antibody [mAbcam 3609] (ab3609)
    Western blot - Anti-Aurora B antibody [mAbcam 3609] (ab3609)
    All lanes : Anti-Aurora B antibody [mAbcam 3609] (ab3609) at 1 µg/ml

    Lane 1 : Hek293 Whole Cell Lysate
    Lane 2 : Jurkat Whole Cell Lysate
    Lane 3 : HeLa Whole Cell Lysate
    Lane 4 : HeLa Whole Cell Lysate- Nocodozole Stimulated

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Absorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [mAbcam 3609] (ab3609)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [mAbcam 3609] (ab3609)

    SKN-SH cells were fixed in 4% paraformaldehyde for 10 mins, permeabilized in PBS-0.5% Triton X-100 for 5 mins and incubated for 30 minutes with ab3609 (1/50 dilution). The slides were blocked with 4mg/ml BSA. The slides were rinsed once in PBS-Triton (0.1%), twice in PBS then incubated with the secondary antibody for 30 mins. The DNA is stained with DAPI (blue). Staining with ab3609 can be seen in green.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [mAbcam 3609] (ab3609)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [mAbcam 3609] (ab3609)Image courtesy of Human Protein Atlas
    Image courtesy of Human Protein Atlas

    Paraffin embedded human liver tumor tissue was incubated with ab3609 (1/35 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab3609 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [mAbcam 3609] (ab3609)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [mAbcam 3609] (ab3609)Image courtesy of an abreview submitted by Dr. Kirk McManus, University of Manitoba/MICB, Canada
    ab3609 (1/200) staining Aurora B in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus/ chromosomes (red). For further experimental details please refer to Abreview.

    See Abreview

参考文献 (18)

ab3609 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab3609 は 18 報の論文で使用されています。

  • Velásquez ZD  et al. Toxoplasma gondii-induced host cellular cell cycle dysregulation is linked to chromosome missegregation and cytokinesis failure in primary endothelial host cells. Sci Rep 9:12496 (2019). PubMed: 31467333
  • Liu S  et al. Nuclear envelope assembly defects link mitotic errors to chromothripsis. Nature 561:551-555 (2018). PubMed: 30232450
  • Park J  et al. USP35 regulates mitotic progression by modulating the stability of Aurora B. Nat Commun 9:688 (2018). PubMed: 29449677
  • Gupta DK  et al. Tension-induced cytokinetic abscission in human fibroblasts. Oncotarget 9:8999-9009 (2018). PubMed: 29507669
  • Noguchi K  et al. Functional Effects of AKT3 on Aurora Kinase Inhibitor-induced Aneuploidy. J Biol Chem 292:1910-1924 (2017). PubMed: 28028179
View all Publications for this product

レビューと Q&A

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1-6 of 6 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Aurora B antibody [mAbcam 3609]

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Heart)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate buffer
Permeabilization
Yes - 0.025% Triton X-100
Specification
Heart
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

投稿 Feb 05 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Aurora B antibody [mAbcam 3609]

 Inconclusive
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (3T3-L1 fibroblast)
Permeabilization
Yes - Triton X100
Specification
3T3-L1 fibroblast
Blocking step
0.1% donkey serum and 0.2% fish skin gelatin as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 20°C
Fixative
Formaldehyde
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Abcam user community

Verified customer

投稿 Dec 06 2016

Question

dear correspondent,

We have lately purchased several Abs from Abcam and i have a question related to the purification method.
In the datasheets of several monoclonal antibodies is mentioned that they are purified as 'IgG fraction'. Could you exlain to me what that means? Is a anti-IgG column used or an protein A/G column (or any other technique)? This is very important information to us, as we need to consider protein A/G contamination to explain data obtained in our R&D dept. in experiments using these antibodies.

More specifically, the question relates to the following antibodies (data sheets of all those abs mention 'IgG fraction' in 'purity' annotation):

ab3609-100 (anti-Aurora B)
ab51841-100 (anti-TATA binding protein)
ab37373 (goat control IgG)
ab46540 (rabbit control IgG)


Thanks in advance and hope to hear your answer soon,

Read More
Answer

Thank you for your enquiry.

I have confirmed the purification details of the antibodies you listed and have updated the datasheets where necessary:

ab3609 (Aurora B)
Protein G purified

ab51841 (TATA binding protein)
Protein G purified

ab37373 (goat control IgG)
Proprietary combination of ammonium sulfate precipitation, ion exchange, and/or other chromatography techniques.

ab46540 (rabbit control IgG)
Affinity purified against the antigen.

I hope this will be helpful. If you require any further information on the purification of other products, or any other questions, please do not hesitate to contact me.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Aurora B antibody [mAbcam 3609]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TritonX100 in PBS
Read More

Dr. Kirk Mcmanus

Verified customer

投稿 Mar 30 2010

Western blot abreview for Anti-Aurora B antibody [mAbcam 3609]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293# HeLa(+)# NIH-3T3# PC12#9L)
Loading amount
30 µg
Specification
HEK293# HeLa(+)# NIH-3T3# PC12#9L
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Read More

Mr. Antibody Solutions

Verified customer

投稿 Aug 26 2008

Question

BATCH NUMBER 123472 ORDER NUMBER 20600471 DESCRIPTION OF THE PROBLEM No signal SAMPLE Human cell lines, CAHPV10, Hela, Du145, whole protein extract. PRIMARY ANTIBODY ab3609, 1 hr in 0.5% milk DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED ab9484 as a loading control, numerous other primaries on the same blots. ANTIBODY STORAGE CONDITIONS Aliquoted and stored at -20C upon receipt. SAMPLE PREPARATION RIPA+PI's AMOUNT OF PROTEIN LOADED 20ug ELECTROPHORESIS/GEL CONDITIONS non-reducing, 12% gel TRANSFER AND BLOCKING CONDITIONS 5% milk in TBS-T, 1 hr. SECONDARY ANTIBODY anti-mouse HRP from Amersham, 1hr in 0.5% milk HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Varied primary antibody concentration between 1:250 and 1:1000 (recommended). Have tried on different cell lines, with and without other antibodies present. ADDITIONAL NOTES GAPDH loading control and all other antibodies tested work great using my current protocol. I have yet to see any band for AurB using this antibody.

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Answer

Thank you for contacting us for technical support with ab3609. I'm sorry to hear you are experiencing problems with this antibody. I would expect it to work in HeLa lysate as we have tested the antibody in this lysate and got a nice band. I think the problem could be due to the following: -non reducing conditions. These have not been tried and may not be suitable for this antibody, therefore try reducing conditions - blocking: sometimes the milk prevents the antibody from binding. I would recommend to block in 1% BSA -not enough antibody: try to incubate overnight at 4C in TBST A few more tips to improve signals are to use an Ecl+ or Pierce Super Signal detection kit rather than Ecl;please also make sure that you have plenty of fresh protease inhibitors; check that your secondary works with other primary antibodies. If you still experience problems with reducing conditions, a changed in blocking and an overnight incubation please do not hesitate to contact me and I can offer you a replacement vial or refund as you purchased the antibody in September. Best of luck with your next experiments,

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