Overview

  • Product name
  • Description
    Rabbit polyclonal to Bub3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, EMSAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide derived from residues 300 to the C-terminus of Human Bub3.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • HeLa Nuclear Extract;

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS. pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab4180 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 1/500. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
EMSA Use at an assay dependent concentration. PubMed: 22210889

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

  • Anti-Bub3 antibody (ab4180) at 1/500 dilution + HeLa Nuclear Extract at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution

    Predicted band size: 37 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Western blot of ab4180 on 20ug HeLa Nuclear Extract at 1/500.

    Secondary ab: Goat anti-rabbit IgG HRP conjugate ab6721 (1/2000 - 1 min exposure)

References

This product has been referenced in:
  • Zhu S  et al. A motif from Lys216 to Lys222 in human BUB3 protein is a nuclear localization signal and critical for BUB3 function in mitotic checkpoint. J Biol Chem 290:11282-92 (2015). Read more (PubMed: 25814666) »
  • Burns SS  et al. Histone deacetylase inhibitor AR-42 differentially affects cell-cycle transit in meningeal and meningioma cells, potently inhibiting NF2-deficient meningioma growth. Cancer Res 73:792-803 (2013). WB . Read more (PubMed: 23151902) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa Cell)
Permeabilization
Yes - 0.5% Trition-X
Specification
HeLa Cell
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Hazheen Shirnekhi

Verified customer

Submitted Mar 01 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293)
Loading amount
10 µg
Specification
293
Gel Running Conditions
Reduced Denaturing (12.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 31 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Loading amount
10 µg
Specification
HeLa cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Mar 18 2009

Answer

Thank you for your enquiry. Having performed homology searches of the immunogens of our Bub3 antibodies with the Drosophila protein I regret to inform you that neither antibody seem suitable for this species due to low sequence homology. I'm sorry I cannot help you more in this occasion, please do not hesitate to contact me if I can be of further assistance.

Read More

Answer

Thank you for your enquiry. This should clear things up for you - ab4180 is the antibody (Rabbit polyclonal to Bub3) and ab13739 is the blocking peptide that can be used with ab4180. If you have any additional questions, please contact us again.

Read More

Question
Answer

This antibody has been tested briefly in Immunofluorescence in U20S cells (human). IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees. No staining was observed. However, staining was only attempted once and this is by no means an indication that the antibody doesn't work in IF, it merely did not perform under the conditions above. Please see reviews and the datasheet for details of any further testing.

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

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