Product nameAnti-Bub3 antibody [EPR5319(2)]
See all Bub3 primary antibodies
DescriptionRabbit monoclonal [EPR5319(2)] to Bub3
Tested applicationsSuitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
A synthetic peptide corresponding to residues near the N terminus of Human Bub3 (UniProt ID: O43684)
- WB: Lysates of Jurkat, HeLa, 293T IHC-P: Human breast tissue, Human thyroid carcinoma tissue ICC/IF: 293T cells
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
Immunizing Peptide (Blocking)
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.
|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.|
FunctionHas 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 similaritiesBelongs to the WD repeat BUB3 family.
Contains 5 WD repeats.
Cellular localizationNucleus. 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
- Aa2-050 antibody
- AU019800 antibody
- AU021329 antibody
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.
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
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) »