Product Details

Uniprot No.

Target Names

Alternative Names

Mitotic checkpoint serine/threonine-protein kinase BUB1 beta (EC 2.7.11.1) (MAD3/BUB1-related protein kinase) (hBUBR1) (Mitotic checkpoint kinase MAD3L) (Protein SSK1), BUB1B, BUBR1 MAD3L SSK1

Species Reactivity

Human

Immunogen

A synthesized peptide derived from human BubR1

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

7H4

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
FC	1:20-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

BUB1B, also known as BubR1, serves as a critical mitotic checkpoint kinase that ensures accurate chromosome segregation during cell division. This serine/threonine kinase functions as a key component of the spindle assembly checkpoint, preventing premature anaphase onset until all chromosomes achieve proper bipolar attachment. Dysregulation of BUB1B has been implicated in chromosomal instability and aneuploidy, making it a significant target in cancer research and studies of cell cycle control mechanisms.

This recombinant monoclonal antibody, clone 7H4, offers the reproducibility and consistency that demanding research applications require. Generated against a synthetic peptide derived from human BubR1 and produced using recombinant technology, this antibody provides sequence-defined specificity with minimal lot-to-lot variation, ensuring your experimental results remain comparable across extended studies.

Validation by western blot demonstrates reliable detection of BUB1B in human K562 and U-251 cell lysates, with an observed band at approximately 125 kDa. The slight difference from the predicted molecular weights of 106-122 kDa likely reflects post-translational modifications, including phosphorylation events characteristic of this active kinase. Flow cytometry validation in HeLa cells confirms the antibody's utility for single-cell analysis, with clear separation from isotype control, enabling quantitative assessment of BUB1B expression levels across cell populations.

The flexibility to use this antibody across western blotting, flow cytometry, and ELISA applications supports diverse experimental workflows, from protein expression profiling to functional studies. Researchers investigating mitotic checkpoint signaling, chromosomal instability in cancer models, or fundamental cell cycle regulation will find this antibody a dependable tool for advancing their work.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Target Background

Function

Essential component of the mitotic checkpoint. Required for normal mitosis progression. The mitotic checkpoint delays anaphase until all chromosomes are properly attached to the mitotic spindle. One of its checkpoint functions is to inhibit the activity of the anaphase-promoting complex/cyclosome (APC/C) by blocking the binding of CDC20 to APC/C, independently of its kinase activity. The other is to monitor kinetochore activities that depend on the kinetochore motor CENPE. Required for kinetochore localization of CENPE. Negatively regulates PLK1 activity in interphase cells and suppresses centrosome amplification. Also implicated in triggering apoptosis in polyploid cells that exit aberrantly from mitotic arrest. May play a role for tumor suppression.

Gene References into Functions

Human gastric cancer tissues with low BUBR1 expression showed no eNOS expression. A decrease in BUBR1 reduced eNOS bioavailability through a pathway other than eNOS phosphorylation. PMID: 30396924
Authors show that two distinct pools of BubR1/Bub3 exist at kinetochores and we uncouple these with defined BubR1/Bub3 mutants to address their function. PMID: 27457023
we found that FOXM1 inhibitor attenuated tumorigenesis and radioresistance of glioblastoma (GBM) both in vitro and in vivo. Altogether, BUB1B promotes tumor proliferation and induces radioresistance in GBM, indicating that BUB1B could be a potential therapeutic target for GBM. PMID: 29039578
Results from phylogenomic study identified of a novel conserved cassette of short linear motifs in BubR1 essential for the spindle checkpoint PMID: 28003474
BubR1 N-terminal domain was necessary, but not sufficient to protect against aneuploidy and cancer. In contrast, BubR1 lacking the internal Cdc20-binding domain provided protection against both, which coincided with improved microtubule-kinetochore attachment error correction and spindle assembly checkpoint activity. PMID: 27528194
Low BUB1B expression is associated with Chromophobe Renal Cell Carcinomas. PMID: 28807937
Whether carriers of pathogenic BUB1B mutations, such as the parents of MVA syndrome patients, have an increased risk for cancer remains of interest, as studies in mice have suggested that haploinsufficiency of BUB1B may cause an increase in carcinogen-induced tumors PMID: 27239782
structure of the PP2A B56-BubR1 complex provides important insights into how the B56 subunit directs the recruitment of PP2A to specific targets. PMID: 27350047
Overexpression of BUB1B is associated with Invasive Breast Cancer. PMID: 27165245
we showed that BubR1 and Mad2 are overexpressed in oral squamous cell carcinoma cell lines and linked such overexpression to attenuated spindle assembly checkpoint activity. We also showed BubR1 overexpression to be associated with advanced stage and tumour size. PMID: 25754611
The integrity of the mitotic checkpoint complex depends on the specific recognition between BubR1 and Bub3, for which the BubR1 Gle2 binding sequence motif is essential. PMID: 27030009
We show that kinetochore recruitment of BUBR1 and BUB3 by BUB1 is dispensable for SAC activation PMID: 26148513
Data suggest that both BubR1 and SNCG may be promising predictive marker rather than prognostic marker in patients with breast cancer. PMID: 26191236
BubR1 knockdown significantly decreased cellular invasion but slightly affect cellular proliferation on both Ca9-22 and Cal-27 cells. PMID: 26151845
BubR1 contributes to preventing premature aging. [review] PMID: 25964054
Suggest human papillomavirus E2 protein provokes BUBR1-dependent aneuploidy in HPV-induced cervical cancer. PMID: 25789401
In conclusion, the results presented here suggest that Mad2 and BubR1 could be used as prognostic markers of tumor progression and new pharmacological targets in the treatment for gastric cancer . PMID: 25483095
Co-depletion of MAD2 and BUBR1 causes cell cycle arrest and cell death in addition to aneuploidy. PMID: 24687487
By sequestering PIDD at the kinetochore, BubR1 acts to delay PIDDosome formation until the next cycle, defining a new mechanism by which cells evade apoptosis during mitosis. PMID: 25936804
a Cdc20 binding site in BubR1 facilitates both spindle assembly checkpoint signalling and silencing PMID: 25482201
study shows that SIRT2 is a deacetylase for BubR1 K250, although the abnormally prolonged SAC activation observed in SIRT2 knockdown cells is not accompanied by a change in BubR1 levels or by delayed progression from prometaphase to anaphase PMID: 25285631
YAP constitutively associated with BubR1 (BUB1-related protein kinase), and knockdown of BubR1 relieved YAP-driven hyperactivation of the spindle checkpoint. PMID: 25605730
Data indicate that cell cycle protein Bub3-mediated kinetochore recruitment of BubR1 kinase enhances mitotic checkpoint signaling. PMID: 25246557
the BubR1M-Cdc20 interaction indirectly contributes to mitotic checkpoint complex homeostasis PMID: 25505175
The ABBA motif in cyclin A is required for its proper degradation in prometaphase through competing with BUBR1 for the same site on CDC20 PMID: 25669885
Our findings indicated an interplay between BUBR1 and p53 in colorectal cancer. Altered expression of both molecules was associated with chromosomal instability. PMID: 25275037
the loss of BubR1 levels with age is due to a decline in NAD(+) and the ability of SIRT2 to maintain lysine-668 of BubR1 in a deacetylated state, which is counteracted by the acetyltransferase CBP. PMID: 24825348
In acute myeloid leukemia, repression of BubR1 is associated with enhanced anaphase-promoting complex activity. PMID: 23812934
p53 deficiency may lead to the failure of BubR1 downregulation by OS and that p53 deficiency and BubR1 accumulation could contribute to gastric carcinogenesis associated with aneuploidy. PMID: 24156017
BubR1 overexpression was associated with cell proliferation and may play a role in the carcinogenesis of gastrointestinal diffuse large B cell lymphoma PMID: 23400934
These results suggest that the malignant transformation of plexiform neurofibroma is associated with distinct changes in the expression of BUB1B, PBK and NEK2 PMID: 23370767
High levels of BubR1 were less sensitive to the anti-microtubule drugs paclitaxel and nocodazole in esophageal squamous cell carcinoma. PMID: 23128493
Data suggest that BubR1 counteracts Aurora B kinase activity at improperly attached kinetochores by recruiting B56-PP2A phosphatase complexes. PMID: 23345399
Reduced BubR1 expression is strongly associated with longer survival in prostate cancer patients. PMID: 23475578
Mad2 Binding Induces a Functional Switch in Cdc20,Enabling BubR1 Binding. PMID: 23791783
findings highlight the insufficiency of BUB1 haploinsufficiency to directly stimulate tumourigenesis, and suggest that other factors may be more critical to this process. PMID: 23440991
Rsf-1 increases the frequency of abnormal mitotic events by disrupting hBubR1-Cdc20 interactions. PMID: 23536579
Results suggest that targeting the GLEBS domain activity of BUB1B may provide a therapeutic window for glioblastoma. PMID: 23154965
Elevated BUBR1 expression was associated with poor survival in early stage breast cancer patients PMID: 23392733
The PLK1 and BUBR1 cooperate to stabilize kinetochore-microtubule interactions by regulating PP2A-B56alpha-mediated dephosphorylation of Aurora B substrates at the kinetochore-microtubule interface. PMID: 23079597
It was shown that the state of CENP-E-dependent BubR1 autophosphorylation in response to spindle microtubule capture by CENP-E is important for kinetochore function in achieving accurate chromosome segregation. PMID: 22801780
Results reveal that BubR1 sumoylation plays an important role in its timely removal from the kinetochores and the checkpoint inactivation, thus allowing normal anaphase entry and chromosome segregation. PMID: 22374677
It was shown that Mad3/BUBR1 and BUB1 paralogous pairs arose by nine independent gene duplications throughout evolution. It was also shown that putative catalysis by human BUBR1 is dispensable for error-free chromosome segregation. PMID: 22698286
p31(comet) negatively regulates the spindle assembly checkpoint by extracting Mad2 from the MCC. PMID: 22100920
a new type of post-translational modification that is essential for BubR1 function during mitosis. PMID: 22167194
we identify the Blinkin motif critical for interaction with BUBR1, define the stoichiometry and affinity of the interaction, and present a 2.2 Angstrom resolution crystal structure of the complex PMID: 22000412
DLGAP5-PINK1 and BUB1B-PINK1 were strong predictors of disease-free survival and overall survival, respectively, among adult patients with ACT. PMID: 22048964
Aging-related loss of BubR1 and subsequent impairment of reactivity to reactive oxygen species may explain reduced proliferative capacity of aged smooth muscle cells. PMID: 21550059
Our data imply the possibility that BUBR1 may be involved in the progression of oral squamous cell carcinoma, and suggest that BUBR1 may be a promising prognostic marker in patients with OSCC PMID: 21069850
BUBR1 and closed MAD2 (C-MAD2) interact directly to assemble a functional mitotic checkpoint complex PMID: 21525009

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Involvement in disease

Premature chromatid separation trait (PCS); Mosaic variegated aneuploidy syndrome 1 (MVA1)

Subcellular Location

Cytoplasm. Nucleus. Chromosome, centromere, kinetochore. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Note=Cytoplasmic in interphase cells. Associates with the kinetochores in early prophase. Kinetochore localization requires BUB1, PLK1 and KNL1.

Protein Families

Protein kinase superfamily, Ser/Thr protein kinase family, BUB1 subfamily

Tissue Specificity

Highly expressed in thymus followed by spleen. Preferentially expressed in tissues with a high mitotic index.

Database Links

HGNC: 1149

OMIM: 176430

KEGG: hsa:701

STRING: 9606.ENSP00000287598

UniGene: Hs.513645

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Code	CSB-RA263021A0HU
Size	US$210
	Order now
Image	Western Blot Positive WB detected in: K562 whole cell lysate, U-251 whole cell lysate All lanes: BubR1 antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 120, 106, 122 kDa Observed band size: 125 kDa Overlay histogram showing Hela cells stained with CSB-RA263021A0HU (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followedby the antibody (1µg/110⁶ cells) for 1 h at 4℃.The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4℃. Control antibody (green line) was Rabbit IgG (1µg/110⁶ cells) used under the same conditions. Acquisition of >10,000 events was performed.
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BUB1B Recombinant Monoclonal Antibody

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