Overview

  • Product name

  • Description

    Mouse monoclonal to Bub1
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • General notes

    This product was changed from ascites to tissue culture supernatant on 25/02/19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: None
    PBS, pH 7.2
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

Applications

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.

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 122 kDa.
ICC/IF Use at an assay dependent concentration.

Use with methanol fixed cells.

Flow Cyt Use at an assay dependent concentration.

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

Target

  • Function

    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.
  • Tissue specificity

    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.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
    Contains 1 BUB1 N-terminal domain.
    Contains 1 protein kinase domain.
  • Domain

    The KEN box is required for its ubiquitination and degradation.
    BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
  • Post-translational
    modifications

    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.
  • Cellular localization

    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
  • Database links

  • Alternative names

    • 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

Images

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

    Predicted band size: 122 kDa



    This image was generated using the ascites version of the product.

  • 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.

    This image was generated using the ascites version of the product.

    See Abreview

  • 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.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:

  • Alfonso-Pérez T  et al. MAD1-dependent recruitment of CDK1-CCNB1 to kinetochores promotes spindle checkpoint signaling. J Cell Biol 218:1108-1117 (2019). Read more (PubMed: 30674583) »
  • Hayward D  et al. CDK1-CCNB1 creates a spindle checkpoint-permissive state by enabling MPS1 kinetochore localization. J Cell Biol 218:1182-1199 (2019). Read more (PubMed: 30674582) »
See all 20 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No

Dr. Kirk Mcmanus

Verified customer

Submitted May 08 2008

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