Overview

  • Product name

    Anti-Bub3 antibody [EPR5319(2)]
    See all Bub3 primary antibodies
  • Description

    Rabbit monoclonal [EPR5319(2)] to Bub3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues near the N terminus of Human Bub3 (UniProt ID: O43684)

  • Positive control

    • WB: Lysates of Jurkat, HeLa, 293T IHC-P: Human breast tissue, Human thyroid carcinoma tissue ICC/IF: 293T 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR5319(2)
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab133699 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 1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).Can be blocked with Human Bub3 peptide (ab226013).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/250 - 1/2000.

Target

  • Function

    Has a dual function in spindle-assembly checkpoint signaling and in promoting the establishment of correct kinetochore-microtubule (K-MT) attachments. Promotes the formation of stable end-on bipolar attachments. Necessary for kinetochore localization of BUB1. Regulates chromosome segregation during oocyte meiosis. The BUB1/BUB3 complex plays a role in the inhibition of anaphase-promoting complex or cyclosome (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.
  • Sequence similarities

    Belongs to the WD repeat BUB3 family.
    Contains 5 WD repeats.
  • Cellular localization

    Nucleus. Chromosome > centromere > kinetochore. Starts to localize at kinetochores in prometaphase I (Pro-MI) stage and maintains the localization until the metaphase I-anaphase I (MI-AI) transition.
  • Information by UniProt
  • Database links

  • Alternative names

    • Aa2-050 antibody
    • AU019800 antibody
    • AU021329 antibody
    • AU043350 antibody
    • AW146323 antibody
    • BUB 3 antibody
    • BUB 3L antibody
    • BUB3 (budding uninhibited by benzimidazoles 3, yeast) homolog antibody
    • bub3 antibody
    • BUB3 budding uninhibited by benzimidazoles 3 homolog antibody
    • BUB3 mitotic checkpoint protein antibody
    • BUB3_HUMAN antibody
    • BUB3L antibody
    • Budding uninhibited by benomyl antibody
    • Budding uninhibited by benzimidazoles 3 antibody
    • Budding uninhibited by benzimidazoles 3 homolog (S. cerevisiae) antibody
    • Budding uninhibited by benzimidazoles 3 homolog (yeast) antibody
    • Budding uninhibited by benzimidazoles 3 homolog antibody
    • Budding uninhibited by benzimidazoles 3, S. cerevisiae, homolog of antibody
    • C78067 antibody
    • hBUB3 antibody
    • Mitotic checkpoint component antibody
    • Mitotic checkpoint protein bub3 antibody
    • OTTHUMP00000020679 antibody
    • OTTHUMP00000020680 antibody
    • WD repeat type I transmembrane protein A72.5 antibody
    see all

Images

  • All lanes : Anti-Bub3 antibody [EPR5319(2)] (ab133699) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution

    Predicted band size: 37 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?

  • Immunocytochemistry/Immunofluorescence analysis of HepG2 (human hepatocellular carcinoma) labelling Bub3 with purified ab133699 at 1/2000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • Immunohistochemical analysis of paraffin-embedded human breast tissue labelling Bub3 using ab133699 at 1/100 dilution
  • Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue labelling Bub3 using ab133699 at 1/100 dilution
  • Immunofluorescent staining Bub3 in 293T cells using ab133699 at 1/250 dilution

References

This product has been referenced in:

  • Duggal S  et al. Defining the Akt1 interactome and its role in regulating the cell cycle. Sci Rep 8:1303 (2018). Read more (PubMed: 29358593) »
  • Kleyman M  et al. STAG2 promotes error correction in mitosis by regulating kinetochore-microtubule attachments. J Cell Sci 127:4225-33 (2014). ICC/IF ; Human . Read more (PubMed: 25074805) »
See all 2 Publications for this product

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