BUB3 Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA549049A0HU
Size	US$210
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Image	IHC image of CSB-RA549049A0HU diluted at 1:100 and staining in paraffin-embedded human glioma cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-RA549049A0HU diluted at 1:100 and staining in paraffin-embedded human testis tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Immunofluorescence staining of HepG2 cell with CSB-RA549049A0HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). Overlay Peak curve showing Hela cells stained with CSB-RA549049A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Target Background

Product Details

Uniprot No.

O43684

Target Names

BUB3

Alternative Names

Aa2-050 antibody; AU019800 antibody; AU021329 antibody; AU043350 antibody; AW146323 antibody; BUB 3 antibody; BUB 3L antibody; BUB3 (budding uninhibited by benzimidazoles 3; yeast) homolog antibody; bub3 antibody; BUB3 budding uninhibited by benzimidazoles 3 homolog antibody; BUB3 mitotic checkpoint protein antibody; BUB3_HUMAN antibody; BUB3L antibody; Budding uninhibited by benomyl antibody; Budding uninhibited by benzimidazoles 3 antibody; Budding uninhibited by benzimidazoles 3 homolog (S. cerevisiae) antibody; Budding uninhibited by benzimidazoles 3 homolog (yeast) antibody; Budding uninhibited by benzimidazoles 3 homolog antibody; Budding uninhibited by benzimidazoles 3; S. cerevisiae; homolog of antibody; C78067 antibody; hBUB3 antibody; Mitotic checkpoint component antibody; Mitotic checkpoint protein bub3 antibody; OTTHUMP00000020679 antibody; OTTHUMP00000020680 antibody; WD repeat type I transmembrane protein A72.5 antibody

Species Reactivity

Human

Immunogen

A synthesized peptide derived from human BUB3

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

3E5

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, IHC, IF, FC

Recommended Dilution

Application	Recommended Dilution
IHC	1:50-1:200
IF	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) 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

BUB3 serves as an essential component of the spindle assembly checkpoint, a critical surveillance mechanism that ensures accurate chromosome segregation during cell division. This mitotic checkpoint protein forms complexes with other checkpoint components to monitor kinetochore-microtubule attachments, preventing premature anaphase onset until all chromosomes achieve proper bipolar attachment. Dysregulation of BUB3 and related checkpoint proteins has been implicated in chromosomal instability, a hallmark of many cancers, making this target particularly relevant for researchers investigating cell cycle control, aneuploidy, and tumorigenesis.

This recombinant monoclonal antibody, clone 3E5, offers the reproducibility and consistency that demanding experimental workflows require. Generated through recombinant technology with a defined sequence, this rabbit IgG antibody eliminates the lot-to-lot variability often encountered with traditional hybridoma-derived antibodies, ensuring your results remain comparable across extended studies and collaborative projects.

Validation across multiple platforms demonstrates the antibody's versatility for diverse experimental approaches. Immunohistochemistry studies have confirmed specific staining in paraffin-embedded human glioma cancer tissue and testis tissue at 1:100 dilution using citrate buffer antigen retrieval, revealing expression patterns consistent with BUB3's role in proliferating cell populations. Immunofluorescence analysis in HepG2 cells shows clear cellular localization, while flow cytometry validation in HeLa cells demonstrates robust detection with distinct signal separation from isotype controls.

Whether you are investigating mitotic checkpoint mechanisms, exploring chromosomal instability in cancer models, or characterizing cell cycle dynamics, this antibody provides a reliable tool for studying BUB3 biology across immunohistochemistry, immunofluorescence, and flow cytometry applications in human samples.

Usage

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

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Target Background

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.

Gene References into Functions

Small sequence differences in Bub1 and BubR1 direct Bub3 to different phosphorylated targets in the spindle assembly checkpoint signaling cascade. PMID: 28943088
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
a mutual antagonism exists between Bub3 and dynein to ensure optimal KT-MT attachments PMID: 26526612
the mechanisms of BUB3 in mitotic checkpoint signaling PMID: 25814666
Data show sequential multisite regulation of the microtubule-associated protein KNL1-kinase-adaptor complex BUB1/BUB3 interaction. PMID: 25661489
Data indicate that cell cycle protein Bub3-mediated kinetochore recruitment of BubR1 kinase enhances mitotic checkpoint signaling. PMID: 25246557
Results suggest that these USP7-dependent effects are mediated by decreased levels of spindle assembly checkpoint (SAC) component Bub3. PMID: 25003721
Mutations targeting the Bub3 interface prevent kinetochore recruitment of the SAC kinase Bub1. Crucially, they also cause a checkpoint defect, showing that recognition of phosphorylated targets by Bub3 is required for checkpoint signaling PMID: 24066227
Results showed that neither variants in BUB3 nor variants in MAD2L1 caused any significant effect on the risk of breast cancer PMID: 24711138
Mutations in BUB1 and BUB3 cause mosaic variegated aneuploidy and increase the risk of colorectal cancer at a young age. PMID: 23747338
These results indicate that Bub3 and Cdc20 play additional roles in the integration of cell cycle arrest as transcriptional repressors. PMID: 15388328
Allelic loss at 10q26 in osteosarcoma is reported in the region of BUB3. PMID: 15796961
subpopulation of dynein, containing the DYNLT3 light chain, may be important for chromosome congression, in addition to having a role in the transport of checkpoint proteins from the kinetochore to the spindle pole PMID: 17289665
Using RNA interference and high-resolution microscopy, we analyzed kinetochore-microtubule attachments in Bub3-depleted cells and compared them to those in Bub1- or BubR1-depleted cells. PMID: 18199686
Results suggest a novel molecular mechanism leading to aneuploidy involving interference of TAp73alpha with Bub1 and Bub3 resulting in an altered mitotic checkpoint. PMID: 19182530

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Subcellular Location

Nucleus. Chromosome, centromere, kinetochore.

Protein Families

WD repeat BUB3 family

Database Links

HGNC: 1151

OMIM: 603719

KEGG: hsa:9184

STRING: 9606.ENSP00000357858

UniGene: Hs.418533

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Antibody FAQs What Is an Antibody? How to Choose an Antibody for Scientific Research? The difference between primary antibody and secondary antibod The Meaning of Irregular Antibody Screening Tag Antibodies, Your Good Partner in Protein Research How to Choose the Loading Control Antibodies?

Related pathways

Cell cycle Target Therapy

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301-363-4651 (Available 9 a.m. to 5 p.m. CST from Monday to Friday)

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Address

No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China

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