Recombinant Human CEP63 Protein: Handling & Application Protocol

Recombinant Human CEP63 Protein is a centrosome-associated recombinant protein used in studies of centriole duplication, cell cycle checkpoint regulation, and ciliogenesis, requiring careful reconstitution and storage to maintain full biological activity.

Product Name

Recombinant Human CEP63 Protein

SKU

rp374824

Protein Function

CEP63 (Centrosomal Protein 63) plays a role in centriole duplication and is required for proper cell cycle progression and ciliogenesis.

Recommended Storage

Store lyophilized protein at -20°C or -80°C; avoid repeated freeze-thaw cycles. Once reconstituted, aliquot and store at -80°C for long-term use.

Host/Expression System

Recombinant protein produced in a heterologous expression system; specific host details should be confirmed with product datasheet.

Research Applications

Suitable for use in SDS-PAGE, Western blot, ELISA standard curve generation, protein interaction studies, and cell biology assays related to centrosome function.

Background: CEP63 and Its Biological Significance

CEP63, or Centrosomal Protein 63, is a coiled-coil domain-containing protein that localizes to the centrosome and plays an essential role in centriole duplication during the cell cycle. It functions as a scaffold protein at the centrosome, recruiting other factors necessary for the faithful duplication of centrioles prior to mitosis. Disruption of CEP63 expression or function has been linked to impaired centriole biogenesis, aberrant cell cycle progression, and defects in primary cilia formation, making it a protein of significant interest in cancer biology, developmental biology, and ciliopathy research.

Recombinant Human CEP63 Protein (rp374824) from Aladdin Scientific is produced for use in a variety of biochemical and cell biological research applications, enabling scientists to study CEP63 interactions, develop and validate antibodies, and establish quantitative assays.

Reagent Preparation and Reconstitution

Prior to use, allow the vial of Recombinant Human CEP63 Protein to equilibrate to room temperature for 10–15 minutes. Centrifuge the vial briefly (e.g., 5–10 seconds at low speed) to collect any lyophilized material from the walls and cap before opening.

Reconstitution: Reconstitute the lyophilized protein in sterile deionized water or the buffer recommended on the product datasheet to achieve the desired working concentration. Gently pipette to mix; avoid vigorous vortexing, which may denature the protein. Allow the reconstituted protein to stand for 5–10 minutes at room temperature to ensure complete dissolution.

Aliquoting: Immediately divide the reconstituted protein into single-use aliquots to minimize freeze-thaw cycles. Label each aliquot with the date, concentration, and lot number. Store aliquots at -80°C for long-term use or at -20°C for short-term use (within 1 month). Do not store at 4°C for extended periods, as protein stability will diminish.

Note: Specific reconstitution buffer and final concentration recommendations should be verified against the current product datasheet for SKU rp374824.

Experimental Applications and Usage Guidelines

Recombinant Human CEP63 Protein is suitable for the following research applications:

• SDS-PAGE and Western Blot: Use as a positive control or molecular weight marker for CEP63 detection. Load 50–200 ng per lane as a starting point and optimize as needed.
• ELISA Standard Curve: Serial dilutions of the recombinant protein can be used to construct a standard curve for quantitative CEP63 measurement in biological samples.
• Antibody Validation: Use as an antigen to confirm antibody specificity and sensitivity in immunoassay development workflows.
• Protein-Protein Interaction Studies: Pull-down assays, co-immunoprecipitation controls, or surface plasmon resonance experiments can be performed to characterize CEP63 binding partners.
• Functional Cell Biology Assays: When applicable, the protein may be used in reconstitution experiments to study centrosome assembly in cell-free systems.

Optimal working concentrations should be determined empirically for each assay system. Carrier protein (e.g., 0.1% BSA) may be added to dilution buffers to stabilize the recombinant protein at low concentrations.

Quality Control and Troubleshooting

Quality Control: Aladdin Scientific's Recombinant Human CEP63 Protein is tested for purity by SDS-PAGE and for biological activity where applicable. Refer to the Certificate of Analysis (CoA) supplied with each lot for lot-specific purity, concentration, and QC data.

Common Troubleshooting Issues:

• Protein aggregation: If aggregation is observed after reconstitution, centrifuge the solution at 10,000 × g for 5 minutes at 4°C to remove insoluble material before use. Ensure reconstitution is performed in the correct buffer as specified in the datasheet.
• Low signal in Western blot: Verify antibody compatibility with recombinant protein, optimize antibody concentration, and confirm proper transfer conditions. Check that the protein has not undergone excessive freeze-thaw cycles.
• Degradation: Protein degradation may result from improper storage, repeated freeze-thaw cycles, or protease contamination. Always use protease inhibitors where applicable and maintain cold chain throughout handling.

For technical support, contact Aladdin Scientific's customer service team with the lot number and product SKU (rp374824).

Safety, Disposal, and Regulatory Considerations

Recombinant Human CEP63 Protein is intended for research use only (RUO) and is not approved for diagnostic, therapeutic, or clinical use in humans or animals.

Safety: Handle all recombinant proteins using standard laboratory safety practices. Wear appropriate personal protective equipment (PPE), including gloves and a lab coat. Although this protein is not classified as a hazardous material under standard regulatory frameworks, treat it as a potential allergen and avoid inhalation or skin contact with lyophilized powder.

Disposal: Dispose of protein solutions and contaminated materials in accordance with your institution's biosafety and chemical waste disposal regulations. Inactivate protein solutions with appropriate disinfectant prior to disposal if required by institutional policy.

Export and Regulatory Compliance: This product may be subject to export control regulations depending on the destination country. Confirm compliance with all applicable local, national, and international regulations prior to ordering and use.

Frequently asked questions

How should Recombinant Human CEP63 Protein be reconstituted and stored?

Reconstitute the lyophilized protein in sterile deionized water or the buffer specified on the product datasheet, gently mixing without vortexing. Immediately aliquot the reconstituted protein into single-use volumes and store at -80°C for long-term use. Avoid repeated freeze-thaw cycles, as these can reduce protein activity and promote aggregation.

What are the recommended applications for Recombinant Human CEP63 Protein?

Recombinant Human CEP63 Protein (rp374824) is suitable for use as a positive control in Western blot and SDS-PAGE, as a standard for ELISA-based quantification, for antibody validation, and in protein-protein interaction studies such as pull-down assays. It may also be used in cell-free reconstitution experiments exploring centrosome assembly.

What is the biological role of CEP63, and why is it relevant to my research?

CEP63 is a centrosomal scaffold protein essential for centriole duplication and proper cell cycle progression. Its dysfunction is associated with defects in primary cilia formation (relevant to ciliopathies) and aberrant mitosis (relevant to cancer biology). Researchers studying centrosome biology, cell division, or cilia-related diseases will find CEP63 a valuable research target.

How can I troubleshoot low purity or aggregation of the recombinant protein?

If aggregation is observed, centrifuge the reconstituted solution at 10,000 × g for 5 minutes at 4°C to remove insoluble aggregates before use. Ensure you are using the correct reconstitution buffer and concentration as stated in the datasheet. For purity concerns, verify the CoA provided with the lot and contact Aladdin Scientific technical support with the lot number for further assistance.

References

1. Loffler H, Fechter A, Liu FY, Poppelreuther S, Kramer A. DNA damage-induced centrosome amplification occurs via excessive numbers of progenitor centrioles. Cell Cycle. 2013;12(17):2916–2921.
2. Sir JH, Barr AR, Nicholas AK, et al. A primary microcephaly protein complex forms a ring around parental centrioles. Nat Genet. 2011;43(11):1147–1153.
3. Aladdin Scientific. Recombinant Human CEP63 Protein (rp374824) Product Datasheet. Aladdin Scientific; [current version].

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.