|Application:||IHC-P, IHC-F, ICC, WB|
Data & Images
|Product Name||Anti-Bub3 Antibody|
|Description||Rabbit IgG polyclonal antibody for Mitotic checkpoint protein BUB3(BUB3) detection. Tested with WB, IHC-P, IHC-F, ICC in Human;Mouse;Rat.|
|Cite This Product||Anti-Bub3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1645)|
|Replacement Item||This antibody may replace the following items: sc-28258|sc-22495|sc-19395|sc-376506|sc-19397|sc-136217|sc-22496 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
|Application||IHC-P, IHC-F, ICC, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
|Immunogen||A synthetic peptide corresponding to a sequence in the middle region of human Bub3(76-90aa DHQLKMHDLNTDQEN), identical to the related rat and mouse sequences.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
*carrier free antibody available upon request.
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20˚C for one year. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Mitotic checkpoint protein BUB3|
|Molecular Weight||37155 MW|
|Protein 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.|
|Subcellular 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. .|
|Alternative Names||Mitotic checkpoint protein BUB3;BUB3;|
|Research Areas|||cell biology|cell cycle|cell division|spindle||
Background for Mitotic checkpoint protein BUB3
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Bub3 Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Immunocytochemistry , 0.5-1μg/ml, Human, Mouse, Rat|
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Rat, Human, Mouse
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Bub3 Antibody Images
Click the images to enlarge.
All lanes: Anti Bub3 (PA1645) at 0.5ug/ml
Lane 1: HELA Whole Cell Lysate at 40ug
Lane 2: A549 Whole Cell Lysate at 40ug
Lane 3: JURKAT Whole Cell Lysate at 40ug
Lane 4: COLO320 Whole Cell Lysate at 40ug
Predicted bind size: 37KD
Observed bind size: 37KD
IHC(P): Rat Testis Tissue
IHC(P): Human Mammary Cancer Tissue
ICC: A549 Cell
ICC: HELA Cell
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,