Product nameAnti-Bub3 antibody [EPR5319(2)] (HRP)
See all Bub3 primary antibodies
DescriptionRabbit monoclonal [EPR5319(2)] to Bub3 (HRP)
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Synthetic peptide within Human Bub3 aa 1-100 (N terminal). The exact sequence is proprietary.
- WB: Jurkat, HeLa and HEK 293 whole cell lysates. IHC: Human Duodenum (FFPE)
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. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% Proclin
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab200805 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/5000. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).|
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)] (HRP) (ab200805) at 1/5000 dilution
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab200805 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
IHC image of Bub3 staining in a section of formalin-fixed paraffin-embedded normal human duodenum*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab200805, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab200805 has not yet been referenced specifically in any publications.