Recombinant
RabMAb

Anti-BubR1 antibody [EPR20652] (ab209998)

Overview

  • Product name
    Anti-BubR1 antibody [EPR20652]
    See all BubR1 primary antibodies
  • Description
    Rabbit monoclonal [EPR20652] to BubR1
  • Tested applications
    Suitable for: WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human BubR1 aa 1-250. The exact sequence is proprietary.
    Database link: O60566

  • Positive control
    • WB: Human cell lines HeLa, Raji and HepG2 whole cell lysates. ICC/IF: Human cell lines HeLa, HepG2, A549 and HT-29. IP: Human cell line HeLa whole cell lysate.
  • 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 ab209998 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 1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 119 kDa).
ICC/IF 1/500.
IP 1/30.

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

  • BubR1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209998 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209998 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1000 dilution.


    Lane 1: HeLa whole cell lysate 10 µg (Input).

    Lane 2: ab209998 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209998 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 1 second.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (human colorectal adenocarcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HT-29 cell line.

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

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of A549 cell line.

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

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HepG2 cell line.

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

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody 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 with ab209998 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing subcellular localization of BubR1 changes through mitosis of HeLa cell line (PMID: 25833949). The nuclear counter stain is DAPI (blue).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling BubR1 with ab209998 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.
    -ve control: Secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

  • All lanes : Anti-BubR1 antibody [EPR20652] (ab209998) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 119 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    Positive BubR1 detection by WB is dependent on the number of mitotic cells in the sample and may need optimization for some cell and tissue lysates.

    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: Lane 1 - 10 seconds, lane 2 - 3 seconds, lane 3 - 3 minutes

References

ab209998 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab209998.
Please use the links above to contact us or submit feedback about this product.

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

Sign up