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Product name
Anti-Bub1 antibody
See all Bub1 products (7) ...
Description
Rabbit polyclonal to Bub1
Tested applications
ICC/IF, WBmore details
Cross reactivity
Reacts with
Rat, Human
Immunogen
His-tagged N-terminal bacterially expressed human Bub1 (aa 1-312).
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Purity
Whole antiserum
Clonality
Polyclonal
Isotype
IgG
Research areas
Cancer >> Cell cycle >> Cell division
Cell Biology >> Cell Cycle >> Cell Division >> Spindle
Applications
Show applications key
Our Abpromise guarantee covers the use of ab9000 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notes
ICC/IF: Use at an assay dependent dilution PMID 18199686.
WB: 1/2000. Predicted molecular weight: 122 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
Serine/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 specificity
High 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 similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain.
Domain
The KEN box is required for its ubiquitination and degradation.
BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
Post-translational
modifications
Phosphorylated 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 localization
Nucleus. 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.
Target information above from: UniProt accessionO43683
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Bub1 antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) antibody
see all
Database links
- Entrez Gene: 699 Human
- Entrez Gene: 296137 Rat
- Omim: 602452 Human
- SwissProt: O43683 Human
- Unigene: 469649 Human
Anti-Bub1 antibody images:
Western blot - Bub1 antibody (ab9000)
Predicted band size : 122 kDa
Western blot using ab9000 on Bub1 antigen (lanes 1- 3), and HL-60 whole cell extract.
The Bub1 antigen is only a fragment of full length Bub1 and thus runs at a lower size than the band seen in HL-60 extract.
Immunocytochemistry/ Immunofluorescence - Bub1 antibody (ab9000)
ab9000 staining Bub1 in HCT cells by Immunocytochemistry/ Immunofluorescence. Cells were plated on 12 mm coverslips, pre-extracted with 0.2% Triton X-100 in warm PEM (100 mM PIPES pH6.8, 1mM MgCl2 and 5mM EGTA) for 1 minute, fixed with 3% PFA in PBS, blocked with 3% BSA in PBS for 1 hour, incubated with primary antibody for 16 hrs at 4°C, washed with PBS/0.1% Triton X-100 and incubated with secondary antibody for an additional 1 hour at room temperature. ACA staining in red, Bub1 is green and DNA (DAPI) is in blue.
Image from Sliedrecht T et al, PLoS One. 2010 Apr 22;5(4):e10251, Fig S4.
References for Anti-Bub1 antibody (ab9000)
This product has been referenced in:
- Cui W et al. Roles of MAPK and spindle assembly checkpoint in spontaneous activation and MIII arrest of rat oocytes. PLoS One 7:e32044 (2012). ICC/IF; Rat.Read more (PubMed: 22384134) »
- Sliedrecht T et al. Chemical genetic inhibition of Mps1 in stable human cell lines reveals novel aspects of Mps1 function in mitosis. PLoS One 5:e10251 (2010). ICC/IF; Human.Read more (PubMed: 20422024) »
See all 5 publications for this product
Publishing research using ab9000? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"