Product nameAnti-BubR1 antibody
See all BubR1 primary antibodies
DescriptionSheep polyclonal to BubR1
Tested applicationsSuitable for: IP, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse
Recombinant fragment, corresponding to amino acids 232-603 of Mouse BubR1
- Mitotic NIH/3T3 cells
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.08% Sodium azide
Concentration information loading...
PurityAmmonium Sulphate Precipitation
Our Abpromise guarantee covers the use of ab28193 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 1 - 10 µg/ml.|
|WB||Use a concentration of 1 - 10 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 120 kDa).|
|IHC-P||Use at an assay dependent dilution. PubMed: 20807801|
|ICC/IF||Use at an assay dependent concentration. PubMed: 22024163|
FunctionEssential 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 specificityHighly expressed in thymus followed by spleen. Preferentially expressed in tissues with a high mitotic index.
Involvement in diseaseNote=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 similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain.
DomainThe 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.
modificationsProteolytically 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 localizationCytoplasm. 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
- Beta homolg of S. cerevisiae BUB 1 antibody
- Beta homolg of S. cerevisiae budding uninhibited by benzimidazoles antibody
- BUB 1B antibody
ab28193 staining BubR1 in Mouse metaphase I oocyte by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100. Samples were incubated with primary antibody (1/200) for 15 hours at 4°C. A FITC-conjugated Donkey anti-sheep IgG polyclonal (1/500) was used as the secondary antibody. The signal of BuRI was detected near the chromosome kinetocore of meiosis I stage oocyte.
Anti-BubR1 antibody (ab28193) at 1/3000 dilution + Mouse MII oocytes whole cell lysate at 60 cells
Rabbit Anti-Sheep IgG H&L (HRP) (ab6747) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 120 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes
Mouse embryos were treated briefly with acidic Tyrode’s solution to remove their zona pellucida. They were fixed in 4% paraformaldehyde in PBS and permeabilized in 1% Triton X-100 in PBS. Unspecific binding was blocked with 0.5% Tween-20 in PBS containing 5% bovine serum albumin. Subsequently, the embryos were incubated overnight with ab28193 at a 1/200 dilution. Unspecific binding was blocked with 50% normal goat serum, followed by incubation with an Alexa-Fluor 488 conjugated secondary antibody at a 1/500 dilution.
This product has been referenced in:
- Jiao XF et al. Abce1 orchestrates M-phase entry and cytoskeleton architecture in mouse oocyte. Oncotarget 8:39012-39020 (2017). Read more (PubMed: 28380459) »
- Li X et al. HDAC3 promotes meiotic apparatus assembly in mouse oocytes by modulating tubulin acetylation. Development 144:3789-3797 (2017). Read more (PubMed: 28935703) »