Recombinant Human CEP57 Protein: Handling & Usage Protocol

Recombinant Human CEP57 Protein is a research-grade recombinant protein designed to support investigations into centrosome integrity, microtubule organization, and cell cycle regulation in human cellular systems.

Product Name

Recombinant Human CEP57 Protein

SKU

rp366876

Protein Function

CEP57 is a centrosomal protein involved in microtubule anchoring and centrosome cohesion during the cell cycle

Species Origin

Human (Homo sapiens)

Application Areas

Centrosome biology, cell cycle studies, microtubule dynamics, protein interaction assays

Storage Recommendation

Avoid repeated freeze-thaw cycles; aliquot upon reconstitution and store at -80°C for long-term stability

Background: CEP57 and Its Biological Role

CEP57 (Centrosomal Protein 57) is a conserved centrosomal protein that plays a critical role in maintaining centrosome integrity and regulating microtubule organization during cell division. It is expressed in human somatic cells and is required for proper bipolar spindle formation, ensuring accurate chromosome segregation during mitosis.

Loss-of-function mutations in CEP57 are associated with mosaic variegated aneuploidy (MVA) syndrome, a rare autosomal recessive disorder characterized by constitutional aneuploidies and increased cancer predisposition. Research into CEP57 function therefore has relevance to both fundamental cell biology and translational cancer research.

Recombinant Human CEP57 Protein (rp366876) provides researchers with a well-defined, research-grade tool to study CEP57 protein interactions, biochemical activities, and its role in centrosome-associated pathways in vitro.

Reconstitution and Storage Protocol

Proper reconstitution and storage of Recombinant Human CEP57 Protein are essential to preserve protein activity and experimental reproducibility. Follow the steps below carefully:

1. Centrifugation: Before opening the vial, briefly centrifuge (e.g., 5–10 seconds at low speed) to collect any lyophilized material from the walls and cap of the tube.
2. Reconstitution Buffer: Reconstitute the lyophilized protein in sterile deionized water or an appropriate buffer (e.g., PBS, pH 7.4) as indicated on the product datasheet. Add the reconstitution volume slowly and gently swirl — do not vortex vigorously, as this may denature the protein.
3. Aliquoting: Immediately after reconstitution, divide the protein into single-use aliquots to minimize freeze-thaw cycles.
4. Short-Term Storage: Reconstituted protein may be stored at 4°C for up to 1 week when kept sterile.
5. Long-Term Storage: For extended storage, aliquots should be stored at -80°C. Avoid repeated freeze-thaw cycles, as these will degrade protein integrity and reduce biological activity.
6. Carrier Protein: If the protein will be used at very low concentrations, consider adding a carrier protein (e.g., 0.1% BSA) to prevent adsorption to tube walls.

Note: Always consult the specific lot datasheet supplied with your product for precise reconstitution volumes and buffer recommendations, as conditions may vary by lot.

Experimental Applications

Recombinant Human CEP57 Protein (rp366876) is suitable for a range of in vitro and cell-based research applications, including but not limited to:

• Protein–Protein Interaction Studies: Use CEP57 as bait or prey in pull-down assays or co-immunoprecipitation experiments to identify novel binding partners within the centrosomal protein network.
• Biochemical Characterization: Investigate the microtubule-binding properties of CEP57 using co-sedimentation assays or surface plasmon resonance (SPR) to define binding kinetics.
• ELISA and Western Blot Standards: Use as a positive control or standard for immunodetection-based assays targeting CEP57.
• Antibody Validation: Validate anti-CEP57 antibodies by confirming specific recognition of the recombinant protein in dot blot or Western blot formats.
• Structural Studies: Suitable as a starting material for biophysical characterization including circular dichroism (CD) spectroscopy or limited proteolysis to probe domain organization.

Researchers should optimize assay conditions (protein concentration, buffer composition, incubation time and temperature) empirically for each specific application.

Quality Control and Handling Precautions

To ensure experimental reliability when working with Recombinant Human CEP57 Protein, observe the following quality control practices and handling precautions:

• Purity Verification: Confirm protein purity by SDS-PAGE analysis prior to use. Recombinant proteins are typically supplied at >85% purity as assessed by SDS-PAGE; consult the lot-specific certificate of analysis (CoA) for exact values.
• Endotoxin Levels: For cell-based assays, check the CoA for endotoxin content (LAL assay result). Low endotoxin levels are critical to avoid non-specific cellular responses.
• Activity Testing: Where a functional assay is available (e.g., microtubule co-sedimentation), include a positive control from a validated lot to confirm biological activity of each new lot.
• Avoiding Contamination: Use sterile pipette tips and tubes. Work in a clean environment to prevent proteolytic contamination, which can degrade the recombinant protein.
• Protein Concentration Determination: Measure protein concentration after reconstitution using a Bradford or BCA assay, as the theoretical concentration provided may vary slightly with reconstitution volume.

If the protein shows signs of aggregation (visible turbidity or precipitation), centrifuge at 10,000 × g for 10 minutes at 4°C before use and use only the supernatant.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human CEP57 Protein?

The reconstitution buffer should be selected based on the specific lot datasheet provided with the product (SKU: rp366876). Generally, sterile PBS (pH 7.4) or deionized water is suitable. Always consult the certificate of analysis for lot-specific recommendations.

How should Recombinant Human CEP57 Protein be stored to maintain activity?

After reconstitution, aliquot the protein into single-use volumes and store at -80°C for long-term use. Short-term storage at 4°C is acceptable for up to one week. Repeated freeze-thaw cycles should be strictly avoided, as they compromise protein integrity and biological activity.

What applications is Recombinant Human CEP57 Protein suitable for?

Recombinant Human CEP57 Protein (rp366876) is suitable for protein–protein interaction studies, biochemical characterization of microtubule binding, Western blot and ELISA standards, antibody validation, and biophysical structural studies. Researchers should optimize conditions empirically for each specific assay format.

What is the biological significance of CEP57 in human cells?

CEP57 is a centrosomal protein essential for microtubule anchoring and bipolar spindle formation during mitosis. Mutations in CEP57 are linked to mosaic variegated aneuploidy (MVA) syndrome, highlighting its critical role in chromosome segregation fidelity and genomic stability.

How can I verify the quality of each lot of Recombinant Human CEP57 Protein?

Quality can be assessed by reviewing the lot-specific Certificate of Analysis (CoA), which includes SDS-PAGE purity data and endotoxin levels. For functional verification, include the protein in a validated activity assay (e.g., microtubule co-sedimentation) alongside a previously validated lot as a positive control.

References

1. Snape K, et al. Mutations in CEP57 cause mosaic variegated aneuploidy syndrome. Nature Genetics. 2011.
2. Momotani K, et al. Centrosomal protein CEP57 and its role in microtubule organization. General cell biology reference — specific details require verification.
3. Aladdin Scientific. Recombinant Human CEP57 Protein, SKU rp366876. Product Datasheet.

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