Key features and details
- Rabbit polyclonal to Bub1
- Suitable for: WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Bub1 antibody
See all Bub1 primary antibodies
DescriptionRabbit polyclonal to Bub1
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Human
Recombinant fragment within Human Bub1 (N terminal). The exact sequence is proprietary.
Database link: O43683
- WB: A431 and HEK-293T whole cell lysates. ICC/IF: U-2 OS cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.00
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: 1.21% Tris, 0.75% Glycine, 10% Glycerol (glycerin, glycerine)
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab227228 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/3000. Predicted molecular weight: 122 kDa.|
|ICC/IF||1/100 - 1/1000.|
FunctionSerine/threonine-protein kinase that performs 2 crucial functions during mitosis: it is essential for spindle-assembly checkpoint signaling and for correct chromosome alignment. Has a key role in the assembly of checkpoint proteins at the kinetochore, being required for the subsequent localization of CENPF, BUB1B, CENPE and MAD2L1. Required for the kinetochore localization of PLK1. Plays an important role in defining SGOL1 localization and thereby affects sister chromatid cohesion. Acts as a substrate for anaphase-promoting complex or cyclosome (APC/C) in complex with its activator CDH1 (APC/C-Cdh1). Necessary for ensuring proper chromosome segregation and binding to BUB3 is essential for this function. Can regulate chromosome segregation in a kinetochore-independent manner. Can phosphorylate BUB3. The BUB1-BUB3 complex plays a role in the inhibition of 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. Kinase activity is essential for inhibition of APC/CCDC20 and for chromosome alignment but does not play a major role in the spindle-assembly checkpoint activity. Mediates cell death in response to chromosome missegregation and acts to suppress spontaneous tumorigenesis.
Tissue specificityHigh expression in testis and thymus, less in colon, spleen, lung and small intestine. Expressed in fetal thymus, bone marrow, heart, liver, spleen and thymus. Expression is associated with cells/tissues with a high mitotic index.
Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain.
DomainThe KEN box is required for its ubiquitination and degradation.
BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Upon spindle-assembly checkpoint activation it is hyperphosphorylated and its kinase activity toward CDC20 is stimulated. Phosphorylation at Thr-609 is required for interaction with PLK1, phosphorylation at this site probably creates a binding site for the POLO-box domain of PLK1, thus enhancing the PLK1-BUB1 interaction.
Ubiquitinated and degraded during mitotic exit by APC/C-Cdh1.
Cellular localizationNucleus. Chromosome > centromere > kinetochore. Nuclear in interphase cells. Accumulates gradually during G1 and S phase of the cell cycle, peaks at G2/M, and drops dramatically after mitosis. Localizes to the outer kinetochore. Kinetochore localization is required for normal mitotic timing and checkpoint response to spindle damage and occurs very early in prophase. AURKB, CASC5 and INCENP are required for kinetochore localization.
- Information by UniProt
- Bub1 antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) antibody
Anti-Bub1 antibody (ab227228) at 1/3000 dilution + A431 (human epidermoid carcinoma cell line) whole cell lysate at 30 µg
Predicted band size: 122 kDa
7.5% SDS-PAGE gel.
Anti-Bub1 antibody (ab227228) at 1/1000 dilution + HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg
Predicted band size: 122 kDa
5% SDS-PAGE gel.
Paraformaldehyde-fixed U-2 OS (human bone osteosarcoma epithelial cell line) cells stained for Bub1 (green) using ab227228 at 1/500 dilution in ICC/IF. Counterstain: Alpha Tubulin filaments were labeled with an anti-alpha Tubulin antibody (red).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab227228 has not yet been referenced specifically in any publications.