Anti-CDC27 Monoclonal Antibody for WB, ELISA - P30260

Core Product Specifications and Parameters

Biological Background: CDC27 Function and Localization

• CDC27, also known as APC3 or ANAPC3, is a core subunit of the anaphase promoting complex/cyclosome (APC/C), a large E3 ubiquitin ligase essential for cell cycle progression. Related references: PMID:18485873 PMID:27120157 PMID:27509861
• The APC/C mediates ubiquitination of target proteins, primarily forming Lys-11-linked polyubiquitin chains, but also Lys-48- and Lys-63-linked chains, as well as branched Lys-11/Lys-48 chains, to trigger proteasomal degradation. Related references: PMID:18485873 PMID:29033132
• During the metaphase-to-anaphase transition, APC/C is activated by CDC20, leading to the degradation of cyclin B and securin, thereby driving chromosome segregation and mitotic exit. Related references: PMID:27120157 PMID:27509861
• APC/C activity is tightly regulated by the mitotic checkpoint complex (MCC), which inhibits APC/C until all chromosomes are properly attached to the spindle, preventing premature anaphase onset. Related references: PMID:27509861
• CDC27 localizes to the nucleus, cytoplasm, and mitotic spindle, consistent with its dynamic roles in cell cycle checkpoints and chromosome dynamics.
• The protein contains TPR repeats and is subject to phosphorylation, underscoring its regulatory interactions and post-translational control within the APC/C holoenzyme.
• Alternative splicing generates multiple isoforms, expanding the functional diversity of CDC27 in different cellular contexts.

Experimental Guidance and Technical Tips

• The immunogen comprises amino acids 702–824 of human CDC27, a region likely exposed on the native protein; this design may favor detection of both endogenous and recombinant forms in WB.
• In WB, a band near 92 kDa is expected; confirm specificity using appropriate positive (e.g., HeLa cell lysate) and negative controls, and consider using a loading control.
• For ELISA applications, validate binding against the specific epitope region or full-length CDC27 to ensure assay sensitivity and specificity.
• Cross-reactivity with mouse CDC27 is indicated; test in relevant mouse tissue or cell line samples if targeting murine orthologs.

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