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Immunofluorescence (IF)/Immunocytochemistry (ICC) (Staining HepG2 cells(serum starvation treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab (MBS9612836) and mouse anti-beta tubulin Ab (MBS9610328) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI (blue).)

Rabbit anti-Human, Mouse Bub1 Polyclonal Antibody | anti-Bub1 antibody

Phospho-Bub1 (Thr461) Antibody

Gene Names
BUB1; BUB1A; BUB1L; hBUB1
Reactivity
Human, Mouse
Applications
Immunofluorescence, Immunocytochemistry, ELISA
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Synonyms
Bub1; Polyclonal Antibody; Phospho-Bub1 (Thr461) Antibody; BUB1 budding uninhibited by benzimidazoles 1 homolog; BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast); BUB1 mitotic checkpoint serine/threonine kinase; BUB1; S. cerevisiae; homolog of; BUB1_HUMAN; BUB1A; BUB1L; Budding uninhibited by benzimidazoles 1 (yeast homolog); Budding uninhibited by benzimidazoles 1 homolog; Budding uninhibited by benzimidazoles 1; hBUB1; Homolog of mitotic checkpoint gene BUB1; Mitotic checkpoint gene BUB1; Mitotic checkpoint serine/threonine protein kinase BUB1; Mitotic checkpoint serine/threonine-protein kinase BUB1; Mitotic spindle checkpoint kinase; Putative serine/threonine protein kinase; anti-Bub1 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Phospho-Bub1 (Thr461) Antibody detects endogenous levels of Bub1 only when phosphorylated at Thr461.
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.
Purity/Purification
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Form/Format
Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Concentration
1mg/ml (varies by lot)
Applicable Applications for anti-Bub1 antibody
Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Application Notes
IF/ICC: 1:100-1:500
Peptide ELISA: 1:20,000-1:40,000
Immunogen
A synthesized peptide derived from human Bub1 around the phosphorylation site of Thr461.
Conjugation
Unconjugated
Fragment
Fab fragment
Post Translational Modifications
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.
Subunit Structure
Interacts with BUB3 and KNL1. Interacts (when phosphorylated) with PLK1. The BUB1-BUB3 complex interacts with MAD1L1. (Microbial infection) Interacts with SV40 Large T antigen; this interaction induces activation of a DNA damage response and promotes p53/TP53 stabilization and phosphorylation. (Microbial infection) Interacts with herpes virus 8 protein LANA1.
Similarity
The KEN box is required for its ubiquitination and degradation.BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Subcellular Location
Nucleus. Chromosome>Centromere>Kinetochore.
Note: 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, KNL1 and INCENP are required for kinetochore localization (By similarity).
Preparation and Storage
Store at -20 degree C. Stable for 12 months from date of receipt.

Immunofluorescence (IF)/Immunocytochemistry (ICC)

(Staining HepG2 cells(serum starvation treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab (MBS9612836) and mouse anti-beta tubulin Ab (MBS9610328) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI (blue).)

Immunofluorescence (IF)/Immunocytochemistry (ICC) (Staining HepG2 cells(serum starvation treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab (MBS9612836) and mouse anti-beta tubulin Ab (MBS9610328) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI (blue).)
Related Product Information for anti-Bub1 antibody
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. Required for centromeric enrichment of AUKRB in prometaphase. Plays an important role in defining SGO1 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.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
699
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
122,375 Da
NCBI Official Full Name
mitotic checkpoint serine/threonine-protein kinase BUB1
NCBI Official Synonym Full Names
BUB1 mitotic checkpoint serine/threonine kinase
NCBI Official Symbol
BUB1
NCBI Official Synonym Symbols
BUB1A; BUB1L; hBUB1
NCBI Protein Information
mitotic checkpoint serine/threonine-protein kinase BUB1; mitotic spindle checkpoint kinase; putative serine/threonine-protein kinase; budding uninhibited by benzimidazoles 1 homolog; BUB1 budding uninhibited by benzimidazoles 1 homolog
UniProt Protein Name
Mitotic checkpoint serine/threonine-protein kinase BUB1
UniProt Gene Name
BUB1
UniProt Synonym Gene Names
BUB1L; hBUB1
UniProt Entry Name
BUB1_HUMAN

NCBI Description

This gene encodes a kinase involved in spindle checkpoint function. The kinase functions in part by phosphorylating a member of the miotic checkpoint complex and activating the spindle checkpoint. Mutations in this gene have been associated with aneuploidy and several forms of cancer. [provided by RefSeq, Jul 2008]

Uniprot Description

BUB1: a mitotic-checkpoint protein kinase of the BUB family. Autophosphorylated when the cells enters mitosis. Kinetochore localization is required for normal mitotic timing and checkpoint response to spindle damage. 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. LOF mutations seen in two colon cancer cell lines exhibiting chromosomal instability, in several leukemias and lymphomas, and in a pancreatic cancer cell line. BUB1 functions in mitotic spindle assembly, and may cause chromosomal instability. In mouse BRCA2 knockouts, mutations in BUB1 and BUBR1 are frequently seen in the resulting tumors. BUB1 and BUBR1 are overexpressed, but not mutated, in many gastric cancers.

Protein type: Kinase, protein; EC 2.7.11.1; Protein kinase, Ser/Thr (non-receptor); Protein kinase, Other; Cell cycle regulation; Other group; BUB family

Chromosomal Location of Human Ortholog: 2q14

Cellular Component: nucleoplasm; kinetochore; membrane; cytoplasm; cytosol; condensed nuclear chromosome, pericentric region

Molecular Function: protein serine/threonine kinase activity; protein binding; ATP binding; protein kinase activity

Biological Process: cell proliferation; mitosis; viral reproduction; apoptosis; cell division; mitotic cell cycle checkpoint; mitotic cell cycle spindle assembly checkpoint; regulation of chromosome segregation; regulation of sister chromatid cohesion; mitotic cell cycle; protein amino acid phosphorylation; chromosome segregation

Research Articles on Bub1

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Product Notes

The Bub1 bub1 (Catalog #AAA9612836) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Phospho-Bub1 (Thr461) Antibody reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's Bub1 can be used in a range of immunoassay formats including, but not limited to, Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA). IF/ICC: 1:100-1:500 Peptide ELISA: 1:20,000-1:40,000. Researchers should empirically determine the suitability of the Bub1 bub1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Bub1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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