Recombinant Human CEP250 Protein: Research Guide

Recombinant Human CEP250 Protein is a centrosomal scaffold protein critical for centriole cohesion and ciliogenesis, offered by Aladdin Scientific to support advanced cell biology and ciliopathy research.

Protein Name

Recombinant Human CEP250 (Centrosomal Protein 250)

SKU

rp374831

Biological Function

Structural scaffold at the centrosome; required for centriole-to-centriole cohesion and primary cilia formation

Species Origin

Human (Homo sapiens)

Application Areas

Centrosome biology, ciliogenesis studies, ciliopathy disease modeling, protein interaction assays

Storage Recommendation

Store at -80°C; avoid repeated freeze-thaw cycles (specific buffer conditions subject to datasheet)

Biological Background of CEP250

CEP250, also known as C-Nap1 (Centrosomal NEK2-Associated Protein 1), is a large coiled-coil protein that localizes to the proximal ends of centrioles. It plays an essential structural role in maintaining centriole cohesion during interphase by acting as a linker between paired centrioles. At the onset of mitosis, CEP250 is phosphorylated and displaced by the kinase NEK2, triggering centrosome separation and bipolar spindle assembly.

Beyond its role in the cell cycle, CEP250 is implicated in the regulation of primary ciliogenesis. Mutations in the CEP250 gene have been associated with Usher syndrome and other inherited ciliopathies, underscoring its clinical relevance. Recombinant Human CEP250 Protein enables researchers to dissect these molecular mechanisms in vitro through binding assays, structural studies, and functional reconstitution experiments.

Product Specifications and Quality Attributes

Recombinant Human CEP250 Protein (SKU: rp374831) is produced using recombinant expression technology to ensure reproducibility and lot-to-lot consistency. Key quality considerations include:

• Purity: Assessed by SDS-PAGE; specific purity values are provided on the product datasheet and may vary by lot.
• Formulation: Supplied in an appropriate buffer to maintain protein stability; consult the current datasheet for exact buffer composition and concentration.
• Biological Activity: Functional validation data, where available, are reported on the product datasheet.
• Endotoxin Level: Tested and reported per lot; low endotoxin formulations are preferred for cell-based assays.

Researchers should always refer to the current Certificate of Analysis (CoA) accompanying their specific lot for precise specifications.

Recommended Applications

Recombinant Human CEP250 Protein is suitable for a range of in vitro research applications:

• Protein–Protein Interaction Studies: Use as bait or prey in pull-down assays to identify CEP250 binding partners, including NEK2, rootletin, and other centrosomal components.
• Biochemical Assays: Suitable for ELISA-based quantification, western blot standard curves, and kinase phosphorylation assays involving NEK2.
• Structural Biology: Can be used as antigen for antibody production or as a reference standard in structural proteomics workflows.
• Ciliopathy Research: Supports mechanistic studies of Usher syndrome variants and other CEP250-linked ciliopathies by providing a defined recombinant substrate.
• Cell-Free Reconstitution: Enables reconstitution of centrosome cohesion complexes in cell-free systems for mechanistic dissection.

Always validate the protein for your specific assay format before committing to large-scale experiments.

Handling, Storage, and Stability Guidelines

Proper handling of Recombinant Human CEP250 Protein is essential to preserve its integrity and activity:

• Storage Temperature: Store at -80°C for long-term preservation. For short-term use (up to one week), -20°C may be acceptable — consult the datasheet.
• Aliquoting: Upon first thaw, prepare single-use aliquots to avoid repeated freeze-thaw cycles, which can lead to protein aggregation and loss of activity.
• Thawing: Thaw on ice or at 4°C. Do not heat to accelerate thawing.
• Handling: Avoid vigorous vortexing; gently mix by pipetting. Keep on ice during experimental setup.
• Carrier Protein: If working at very low concentrations, consider supplementing with a carrier protein (e.g., 0.1% BSA) to minimize adsorption losses to tube surfaces.
• Expiry: Check the CoA for shelf life information. Do not use beyond the stated expiry date.

Quality Control and Verification

Aladdin Scientific applies rigorous quality control procedures to each lot of Recombinant Human CEP250 Protein. Standard QC assessments typically include:

• SDS-PAGE Analysis: Confirms apparent molecular weight and purity profile of the recombinant protein.
• Western Blot: Identity confirmation using anti-CEP250 or anti-tag antibodies, as applicable.
• Endotoxin Testing: Performed using LAL (Limulus Amebocyte Lysate) assay or equivalent.
• Concentration Determination: Measured by UV absorbance (A280) or BCA/Bradford assay.

All QC data are documented in the Certificate of Analysis (CoA) shipped with each product. Researchers are encouraged to perform application-specific validation, such as functional activity confirmation, before integrating this protein into critical assay workflows.

Frequently asked questions

What is the biological function of CEP250 and why is it important for research?

CEP250 (C-Nap1) is a centrosomal coiled-coil protein that maintains centriole cohesion during interphase and is displaced by NEK2 kinase at mitotic entry to allow centrosome separation. It is also required for primary ciliogenesis, and mutations in CEP250 are linked to ciliopathies such as Usher syndrome. These roles make it a key target for studying cell cycle regulation, cilia biology, and inherited disease mechanisms.

How should I store Recombinant Human CEP250 Protein to maintain its activity?

Store the protein at -80°C for long-term preservation. Upon first thaw, aliquot into single-use volumes to avoid repeated freeze-thaw cycles. Thaw on ice or at 4°C, and keep samples on ice during use. Always refer to the product datasheet and Certificate of Analysis for lot-specific storage conditions and expiry information.

What applications is Recombinant Human CEP250 Protein (rp374831) validated for?

Recombinant Human CEP250 Protein is suitable for protein–protein interaction studies (pull-down, co-IP), ELISA, western blot standard curves, kinase assays, antigen preparation for antibody generation, and cell-free reconstitution experiments. Application-specific validation data are provided on the product datasheet; users should confirm suitability for their specific experimental format prior to use.

Are there known disease associations with CEP250 that are relevant to translational research?

Yes. Mutations in the CEP250 gene have been associated with Usher syndrome (a condition causing combined hearing and vision loss) and other ciliopathies. Recombinant CEP250 protein can serve as a tool to study pathogenic variants in vitro, evaluate binding partner disruption, and screen potential therapeutic interventions in a biochemical setting.

How do I confirm the identity and purity of the received CEP250 protein lot?

Each lot of Recombinant Human CEP250 Protein is accompanied by a Certificate of Analysis (CoA) that includes SDS-PAGE purity data, concentration, endotoxin level, and identity confirmation results. Researchers can additionally perform an in-house western blot using an anti-CEP250 antibody to independently verify protein identity before use.

References

1. Fry AM, Mayor T, Meraldi P, Stierhof YD, Tanaka K, Nigg EA. C-Nap1, a novel centrosomal coiled-coil protein and candidate substrate of the cell cycle-regulated protein kinase Nek2. J Cell Biol. 1998;141(7):1563-1574. PMID:9647645
2. Graser S, Stierhof YD, Nigg EA. Cep68 and Cep215 (Cdk5rap2) are required for centrosome cohesion. J Cell Sci. 2007;120(Pt 24):4321-4331. PMID:18042621
3. Khateb S, Zelinger L, Mizrahi-Meissonnier L, et al. A homozygous nonsense CEP250 mutation combined with a heterozygous nonsense PCDH15 mutation is associated with Usher syndrome. J Med Genet. 2014;51(7):460-469. PMID:24760770

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