Biological Background of CENPC

Centromere Protein C (CENPC) is a constitutive component of the inner kinetochore, the specialized chromosomal structure that mediates the attachment of chromosomes to spindle microtubules during cell division. CENPC is part of the CCAN (Constitutive Centromere-Associated Network), a multiprotein assembly that bridges centromeric chromatin marked by the histone H3 variant CENP-A and the outer kinetochore KMN network responsible for microtubule capture.

CENPC contains a conserved CENP-C motif and a cupin-like dimerization domain. Through its N-terminal region, CENPC interacts with the Mis12 complex of the outer kinetochore, while its C-terminal region binds directly to CENP-A nucleosomes at centromeric chromatin, thereby serving as a critical linker between chromatin identity and kinetochore function. Loss of CENPC function is associated with chromosome misalignment, mitotic errors, and genomic instability.

Molecular Features of Recombinant Human CENPC Protein

Recombinant Human CENPC Protein (rp364883) is produced using recombinant expression systems to ensure high purity and batch-to-batch consistency suitable for demanding biochemical applications. The recombinant form faithfully reproduces key structural domains of the native human CENPC protein, including the CENP-C motif essential for CENP-A nucleosome recognition and the dimerization domain required for stable kinetochore incorporation.

Because CENPC is a large, intrinsically disordered protein in several of its regions, recombinant production under controlled conditions is important for maintaining the functional integrity of domain interactions. Researchers should verify the specific expression host, tag configuration, and purity grade in the accompanying datasheet (referenced as absea#PP-382) to ensure compatibility with their experimental system.

Research Applications

Recombinant Human CENPC Protein is suited to a range of in vitro and cell-based research applications focused on chromosome biology and mitotic fidelity:

• Kinetochore Reconstitution Assays: CENPC is used to reconstitute inner kinetochore sub-complexes in vitro, enabling mechanistic dissection of how CCAN components are assembled and maintained at centromeres.
• Protein–Protein Interaction Studies: Pull-down, co-immunoprecipitation, and surface plasmon resonance experiments can map CENPC interactions with CENP-A nucleosomes, Mis12 complex subunits, and other CCAN members.
• Structural Biology: Purified recombinant CENPC domains serve as material for X-ray crystallography, cryo-electron microscopy, and NMR studies investigating kinetochore architecture.
• Antibody Validation and ELISA Standards: Recombinant CENPC provides a well-defined antigen for validating anti-CENPC antibodies and generating standard curves in quantitative immunoassays.
• Functional Studies of Mitotic Errors: CENPC can be used in cell-free systems or microinjection experiments to probe consequences of kinetochore disruption on spindle assembly checkpoint signaling.

Storage, Handling, and Quality Considerations

To preserve the activity and structural integrity of Recombinant Human CENPC Protein, researchers should adhere to the storage and handling guidelines provided in the official product datasheet (absea#PP-382). General best practices for recombinant proteins of this class include:

• Store lyophilized or liquid protein at the temperature specified on the datasheet (commonly −20 °C or −80 °C for long-term storage).
• Avoid repeated freeze–thaw cycles, which can cause aggregation and loss of binding activity. Aliquot the protein into single-use volumes upon first reconstitution.
• Reconstitute lyophilized protein in the recommended buffer, allowing the vial to equilibrate at room temperature before gentle mixing.
• Verify protein integrity by SDS-PAGE and activity by a functional assay (e.g., CENP-A nucleosome binding) before use in critical experiments.

Specific details on formulation buffer composition, carrier protein content, endotoxin levels, and purity grade should be confirmed from the product datasheet, as these parameters directly affect experimental outcomes.