Recombinant
RabMAb

Recombinant Anti-BubR1 (phospho S670) antibody [EPR20109] (ab200062)

Overview

  • Product name

    Anti-BubR1 (phospho S670) antibody [EPR20109]
    See all BubR1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20109] to BubR1 (phospho S670)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human BubR1 aa 650-750 (phospho S670). The exact sequence is proprietary.
    Database link: O60566

  • Positive control

    • WB: HeLa whole cell lysate treated with 0.5 µM nocodazole for 24 hours. ICC/IF: HeLa cells.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab200062 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 1/5000. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).
ICC/IF 1/500.

Target

  • Function

    Essential component of the mitotic checkpoint. Required for normal mitosis progression. The mitotic checkpoint delays anaphase until all chromosomes are properly attached to the mitotic spindle. One of its checkpoint functions is to inhibit the activity of the anaphase-promoting complex/cyclosome (APC/C) by blocking the binding of CDC20 to APC/C, independently of its kinase activity. The other is to monitor kinetochore activities that depend on the kinetochore motor CENPE. Required for kinetochore localization of CENPE. Negatively regulates PLK1 activity in interphase cells and suppresses centrosome amplification. Also implicated in triggering apoptosis in polyploid cells that exit aberrantly from mitotic arrest. May play a role for tumor suppression.
  • Tissue specificity

    Highly expressed in thymus followed by spleen. Preferentially expressed in tissues with a high mitotic index.
  • Involvement in disease

    Note=Defects in BUB1B are associated with tumor formation.
    Defects in BUB1B are the cause of premature chromatid separation trait (PCS) [MIM:176430]. PCS consists of separate and splayed chromatids with discernible centromeres and involves all or most chromosomes of a metaphase. It is found in up to 2% of metaphases in cultured lymphocytes from approximately 40% of normal individuals. When PCS is present in 5% or more of cells, it is known as the heterozygous PCS trait and has no obvious phenotypic effect, although some have reported decreased fertility. Inheritance is autosomal dominant.
    Defects in BUB1B are the cause of mosaic variegated aneuploidy syndrome (MVA) [MIM:257300]. MVA is a severe autosomal recessive developmental disorder characterized by mosaic aneuploidies, predominantly trisomies and monosomies, involving multiple different chromosomes and tissues. The proportion of aneuploid cells varies but is usually more than 25% and is substantially greater than in normal individuals. Affected individuals typically present with severe intrauterine growth retardation and microcephaly. Eye anomalies, mild dysmorphism, variable developmental delay, and a broad spectrum of additional congenital abnormalities and medical conditions may also occur. The risk of malignancy is high, with rhabdomyosarcoma, Wilms tumor and leukemia reported in several cases. MVA is caused by biallelic mutations in the BUB1B gene.
  • 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 D-box targets the protein for rapid degradation by ubiquitin-dependent proteolysis during the transition from mitosis to interphase.
    The BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
  • Post-translational
    modifications

    Proteolytically cleaved by caspase-3 in a cell cycle specific manner. The cleavage might be involved in the durability of the cell cycle delay. Caspase-3 cleavage is associated with abrogation of the mitotic checkpoint. The major site of cleavage is at Asp-610.
    Acetylation at Lys-250 regulates its degradation and timing in anaphase entry.
    Ubiquitinated. Degradated by the proteasome.
    Sumoylated by SUMO2 and SUMO3. The sumoylation mediates the association with CENPE at the kinetochore.
    Autophosphorylated in vitro. Intramolecular autophosphorylation is stimulated by CENPE. Phosphorylated during mitosis and hyperphosphorylated in mitotically arrested cells. Phosphorylation at Ser-670 and Ser-1043 occurs at kinetochores upon mitotic entry with dephosphorylation at the onset of anaphase.
  • Cellular localization

    Cytoplasm. Nucleus. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > centrosome. Cytoplasmic in interphase cells. Associates with the kinetochores in early prophase. Kinetochore localization requires BUB1, PLK1 and CASC5.
  • Information by UniProt
  • Database links

  • Alternative names

    • Beta homolg of S. cerevisiae BUB 1 antibody
    • Beta homolg of S. cerevisiae budding uninhibited by benzimidazoles antibody
    • BUB 1B antibody
    • BUB1 budding uninhibited by benzimidazoles 1 homolog beta antibody
    • Bub1A antibody
    • BUB1B antibody
    • BUB1B_HUMAN antibody
    • BUB1beta antibody
    • BUBR1 antibody
    • Budding Uninhibited by Benzimidazoles 1 beta antibody
    • Budding uninhibited by benzimidazoles 1 homolog beta (yeast) antibody
    • hBUBR1 antibody
    • MAD3/BUB1 related protein kinase antibody
    • MAD3/BUB1-related protein kinase antibody
    • MAD3L antibody
    • Mitotic checkpoint gene BUB1B antibody
    • Mitotic checkpoint kinase MAD3L antibody
    • Mitotic checkpoint serine/threonine protein kinase BUB1 beta antibody
    • Mitotic checkpoint serine/threonine-protein kinase BUB1 beta antibody
    • MVA1 antibody
    • OTTHUMP00000160319 antibody
    • Protein SSK1 antibody
    • SSK 1 antibody
    • SSK1 antibody
    see all

Images

  • All lanes : Anti-BubR1 (phospho S670) antibody [EPR20109] (ab200062) at 1/5000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 0.5 µM nocodazole for 24 hours
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 0.5 µM nocodazole for 24 hours, then with alkaline phosphatase for 1 hour

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 120 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling BubR1 (phospho S670) with ab200062 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing strong signal on mitotic phase of HeLa cells, the expression decreased after treatment with lambda protein phosphatase 31°C for 2h.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling BubR1 (phospho S670) with ab200062 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing strong signal on mitotic phase of HeLa cells, the expression decreased after treatment with lambda protein phosphatase 31°C for 2h.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

References

ab200062 has not yet been referenced specifically in any publications.

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