Biological Background of CENPQ

CENPQ (Centromere Protein Q) is a constitutive centromere-associated protein that forms part of the CENP-O complex, which also includes CENP-O, CENP-P, CENP-R, and CENP-U. This complex localizes to the inner kinetochore and plays a critical role in maintaining proper kinetochore architecture and ensuring faithful chromosome segregation during mitosis and meiosis.

CENPQ is a member of the broader CCAN (Constitutive Centromere-Associated Network), a group of proteins that associate with centromeric chromatin throughout the cell cycle. Disruption of CENPQ function has been associated with chromosome misalignment, mitotic errors, and genomic instability, making it a protein of significant interest in cancer biology and cell division research.

Recombinant Human CENPQ Protein produced for research purposes enables investigators to study these interactions in controlled biochemical and cell-based assays without the variability inherent to endogenous protein isolation.

Reconstitution and Storage Protocol

Proper handling of Recombinant Human CENPQ Protein is critical to preserving its activity and structural integrity. Follow the steps below to ensure optimal performance:

• Reconstitution: Briefly centrifuge the vial before opening to consolidate lyophilized material. Reconstitute in an appropriate aqueous buffer (such as PBS, pH 7.4) at the concentration recommended in the product datasheet. Allow the protein to dissolve gently; avoid vigorous vortexing.
• Carrier Protein: If the working concentration is low, consider adding a carrier protein such as 0.1% BSA to prevent adsorption to tube surfaces and maintain stability.
• Aliquoting: Divide the reconstituted protein into single-use aliquots immediately after reconstitution to minimize degradation from repeated freeze-thaw cycles.
• Storage: Store lyophilized protein at -20 °C to -80 °C. Store reconstituted aliquots at -80 °C. Avoid repeated freeze-thaw cycles (limit to fewer than 3).
• Working Conditions: Keep the protein on ice during experimental procedures. Do not leave at room temperature for extended periods.

Note: Always refer to the specific product datasheet (absea#PP-533) supplied with the product for lot-specific reconstitution volumes and buffer recommendations.

Recommended Research Applications

Recombinant Human CENPQ Protein is suitable for multiple research applications in cell biology, biochemistry, and molecular biology:

• Protein–Protein Interaction Studies: CENPQ can be used as bait or prey in pull-down assays or co-immunoprecipitation experiments to characterize interactions within the CENP-O complex and CCAN network.
• In Vitro Binding Assays: Assess direct binding between CENPQ and centromeric DNA, histones, or partner proteins under defined conditions.
• Antibody Development and Validation: Use as an antigen for generating or validating antibodies specific to human CENPQ for Western blot, immunofluorescence, or ELISA applications.
• Structural Biology: Serve as a component in structural studies (e.g., cryo-EM, X-ray crystallography) aimed at resolving kinetochore complex architecture.
• Enzyme Activity and Functional Assays: Investigate post-translational modifications or functional roles in reconstituted kinetochore assembly systems.

All applications should be optimized empirically, as performance may vary depending on experimental conditions, buffer composition, and downstream detection methods.

Quality, Purity, and Regulatory Considerations

Recombinant Human CENPQ Protein from Aladdin Scientific is produced under controlled manufacturing conditions and is intended exclusively for in vitro research use. Key quality considerations include:

• Purity: Recombinant proteins are typically analyzed by SDS-PAGE and/or HPLC to confirm purity. Refer to the Certificate of Analysis (CoA) provided with each lot for purity data specific to your shipment.
• Endotoxin: For cell-based assays where endotoxin contamination may confound results, verify endotoxin levels from the CoA or contact Aladdin Scientific technical support for lot-specific data.
• Identity Confirmation: Protein identity is typically confirmed by mass spectrometry or western blot using anti-CENPQ antibodies. Review the CoA for confirmation methods used.
• Not for Therapeutic Use: This product is supplied for research purposes only. It is not approved, validated, or intended for use in diagnostic, therapeutic, or clinical procedures.
• Lot-to-Lot Variability: As with all biological reagents, researchers should validate each new lot against previous lots in their specific assay system.