BUB3 Antibody

BUB3, also known as Budding Uninhibited by Benzimidazoles 3, is a crucial protein involved in the mitotic checkpoint, a safety mechanism that ensures proper chromosome segregation during cell division. This protein is highly conserved across species, including humans, mice, and yeast. BUB3 contains four WD repeat domains, which are essential for its function in the spindle assembly checkpoint (SAC). The SAC prevents cells from progressing through mitosis until all chromosomes are correctly attached to the spindle apparatus, thereby ensuring accurate chromosome segregation and preventing aneuploidy.

BUB3 functions as a part of the mitotic checkpoint complex (MCC), which also includes BUB1, BUBR1, and MAD2. This complex inhibits the anaphase-promoting complex/cyclosome (APC/C), a ubiquitin ligase that triggers the transition from metaphase to anaphase by targeting specific proteins for degradation. By inhibiting APC/C, BUB3 and its partners delay the onset of anaphase until all chromosomes are properly aligned, thus maintaining genomic stability.

Mouse anti-human BUB3 antibodies are monoclonal or polyclonal antibodies developed in mice to specifically target and bind to the human BUB3 protein. These antibodies are widely used in various scientific applications, including:

• Western Blotting (WB): To detect BUB3 protein levels in different cell and tissue samples.
• Immunohistochemistry (IHC): To visualize BUB3 expression in tissue sections.
• Immunocytochemistry (ICC): To study BUB3 localization within cells.
• Immunoprecipitation (IP): To isolate BUB3 protein complexes from cell lysates.
• Enzyme-Linked Immunosorbent Assay (ELISA): To quantify BUB3 protein levels in various samples.

These antibodies are valuable tools for researchers studying cell cycle regulation, cancer biology, and genomic stability.

Mouse anti-human BUB3 antibodies are developed by immunizing mice with human BUB3 protein or peptides. The immune response generates B cells that produce specific antibodies against BUB3. These B cells are then fused with myeloma cells to create hybridomas, which are screened for the production of high-affinity antibodies. The selected hybridomas are cloned to produce monoclonal antibodies, which are purified and validated for specificity and sensitivity.

Validation of these antibodies involves testing their performance in various applications and species. For example, antibodies are tested for their ability to detect BUB3 in human, mouse, rat, and other species’ samples. Specificity is confirmed by knockdown or knockout experiments, where the absence of BUB3 protein should result in no signal.

Mouse anti-human BUB3 antibodies have been used in numerous studies to investigate the role of BUB3 in cell cycle regulation and cancer. For instance, overexpression or mutations in BUB3 have been linked to various cancers, including colorectal, breast, and lung cancers. By using these antibodies, researchers can study the expression patterns, localization, and interactions of BUB3 in cancer cells, providing insights into its role in tumorigenesis and potential as a therapeutic target.