製品の概要

  • 製品名
  • 製品の詳細
    Mouse monoclonal to Bub1
  • アプリケーション
    適用あり: WB, ICC/IF, Flow Cytmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Recombinant fragment corresponding to Human Bub1 aa 1-130.
    Database link: O43683

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • バッファー
    Preservative: None
    PBS, pH 7.2
  • Concentration information loading...
  • 精製度
    Protein G purified
  • ポリ/モノ
    モノクローナル
  • アイソタイプ
    IgG1
  • 軽鎖の種類
    kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab54893 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 - 5 µg/ml. Predicted molecular weight: 122 kDa.
ICC/IF 1/400. Use with methanol fixed cells.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能
    Serine/threonine-protein kinase that performs 2 crucial functions during mitosis: it is essential for spindle-assembly checkpoint signaling and for correct chromosome alignment. Has a key role in the assembly of checkpoint proteins at the kinetochore, being required for the subsequent localization of CENPF, BUB1B, CENPE and MAD2L1. Required for the kinetochore localization of PLK1. Plays an important role in defining SGOL1 localization and thereby affects sister chromatid cohesion. Acts as a substrate for anaphase-promoting complex or cyclosome (APC/C) in complex with its activator CDH1 (APC/C-Cdh1). Necessary for ensuring proper chromosome segregation and binding to BUB3 is essential for this function. Can regulate chromosome segregation in a kinetochore-independent manner. Can phosphorylate BUB3. The BUB1-BUB3 complex plays a role in the inhibition of APC/C when spindle-assembly checkpoint is activated and inhibits the ubiquitin ligase activity of APC/C by phosphorylating its activator CDC20. This complex can also phosphorylate MAD1L1. Kinase activity is essential for inhibition of APC/CCDC20 and for chromosome alignment but does not play a major role in the spindle-assembly checkpoint activity. Mediates cell death in response to chromosome missegregation and acts to suppress spontaneous tumorigenesis.
  • 組織特異性
    High expression in testis and thymus, less in colon, spleen, lung and small intestine. Expressed in fetal thymus, bone marrow, heart, liver, spleen and thymus. Expression is associated with cells/tissues with a high mitotic index.
  • 配列類似性
    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
    Contains 1 BUB1 N-terminal domain.
    Contains 1 protein kinase domain.
  • ドメイン
    The KEN box is required for its ubiquitination and degradation.
    BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
  • 翻訳後修飾
    Phosphorylated upon DNA damage, probably by ATM or ATR. Upon spindle-assembly checkpoint activation it is hyperphosphorylated and its kinase activity toward CDC20 is stimulated. Phosphorylation at Thr-609 is required for interaction with PLK1, phosphorylation at this site probably creates a binding site for the POLO-box domain of PLK1, thus enhancing the PLK1-BUB1 interaction.
    Ubiquitinated and degraded during mitotic exit by APC/C-Cdh1.
  • 細胞内局在
    Nucleus. Chromosome > centromere > kinetochore. Nuclear in interphase cells. Accumulates gradually during G1 and S phase of the cell cycle, peaks at G2/M, and drops dramatically after mitosis. Localizes to the outer kinetochore. Kinetochore localization is required for normal mitotic timing and checkpoint response to spindle damage and occurs very early in prophase. AURKB, CASC5 and INCENP are required for kinetochore localization.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Bub1 antibody
    • BUB1 budding uninhibited by benzimidazoles 1 homolog antibody
    • BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) antibody
    • BUB1 mitotic checkpoint serine/threonine kinase antibody
    • BUB1, S. cerevisiae, homolog of antibody
    • BUB1_HUMAN antibody
    • BUB1A antibody
    • BUB1L antibody
    • Budding uninhibited by benzimidazoles 1 (yeast homolog) antibody
    • Budding uninhibited by benzimidazoles 1 homolog antibody
    • Budding uninhibited by benzimidazoles 1, S. cerevisiae, homolog of antibody
    • hBUB1 antibody
    • Homolog of mitotic checkpoint gene BUB1 antibody
    • Mitotic checkpoint gene BUB1 antibody
    • Mitotic checkpoint serine/threonine protein kinase BUB1 antibody
    • Mitotic checkpoint serine/threonine-protein kinase BUB1 antibody
    • Mitotic spindle checkpoint kinase antibody
    • Putative serine/threonine protein kinase antibody
    see all

画像

  • ab54893 (1/400) staining Bub1 in Assynchronous Hela cells (green). Cells were fixed in methanol and counterstained with DAPI (red). Please refer to Abreview for further experimental details.

    See Abreview

  • Anti-Bub1 antibody (ab54893) at 2.5 µg/ml + HeLa nuclear Lysate(20µg)

    Predicted band size : 122 kDa
  • Overlay histogram showing HeLa cells stained with ab54893 (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. The cells were then incubated with the antibody (ab54893, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

参考文献

This product has been referenced in:
  • Zhang G  et al. Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling. Nat Commun 8:15822 (2017). IF . Read more (PubMed: 28604727) »
  • Zhang G  et al. Two functionally distinct kinetochore pools of BubR1 ensure accurate chromosome segregation. Nat Commun 7:12256 (2016). WB, IF . Read more (PubMed: 27457023) »

See all 10 Publications for this product

レビューと Q&A

Thank you for your reply.

You should make sure that your samples are reduced and denatured, meaning with ß-mercaptoethanol or DTT and SDS. For the lysis buffer, since this is a nuclear protein, you will need a strong lysis buffer like R...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No
Username

Dr. Kirk Mcmanus

Verified customer

投稿 May 08 2008

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

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