Recombinant Human CFAP418 Protein: Properties & Applications

Recombinant Human CFAP418 Protein is a research-grade recombinant protein derived from the human CFAP418 gene, playing a critical role in cilia assembly and function with emerging relevance to inherited retinal dystrophies.

Protein Name

Recombinant Human CFAP418 Protein

Gene Name

CFAP418 (also known as C8orf37)

Species Origin

Homo sapiens (Human)

Protein Function

Involved in cilia assembly and maintenance; mutations associated with retinal dystrophy and Bardet-Biedl syndrome

Research Applications

Cilia biology, ciliopathy research, retinal disease studies, protein interaction assays

Product SKU

rp373155

Biological Background of CFAP418

CFAP418, also known as C8orf37 (Chromosome 8 Open Reading Frame 37), encodes a cilia- and flagella-associated protein expressed in tissues harboring motile and primary cilia. The protein localizes to the connecting cilium of photoreceptor cells and to the base of cilia in other cell types, where it plays an essential role in cilia assembly and structural maintenance.

Loss-of-function mutations in the CFAP418 gene have been identified in patients with autosomal recessive retinal dystrophy, including cone-rod dystrophy and Bardet-Biedl syndrome (BBS), a ciliopathy characterized by retinal degeneration, obesity, polydactyly, and renal anomalies. These findings underscore the indispensable role of CFAP418 in photoreceptor ciliary integrity and broader ciliary signaling pathways.

As a member of the growing family of ciliopathy-associated proteins, CFAP418 is of significant interest in both basic cilia biology and translational research aimed at understanding and treating inherited retinal degenerative diseases.

Biochemical Properties & Recombinant Protein Characteristics

Recombinant Human CFAP418 Protein is produced using recombinant DNA technology, ensuring a defined and reproducible protein product suitable for a wide range of in vitro research applications. The recombinant protein encompasses the full-length or functionally relevant domain of the human CFAP418 sequence, enabling faithful representation of the native protein's biochemical behavior.

Key characteristics to consider when working with this recombinant protein include its expected molecular weight as predicted from the amino acid sequence, its solubility profile, and its recommended storage conditions—typically at −20°C or −80°C in the presence of a carrier protein such as BSA to minimize freeze-thaw degradation. Researchers should consult the specific product datasheet (SKU: rp373155) for confirmed purity, formulation buffer, and activity data.

The protein is formulated to be compatible with standard assay conditions, including SDS-PAGE, Western blot analysis, ELISA-based binding assays, and co-immunoprecipitation experiments designed to elucidate protein-protein interactions within the ciliary proteome.

Research Applications

Recombinant Human CFAP418 Protein is a versatile reagent for researchers investigating cilia biology, ciliopathies, and inherited retinal diseases. Primary applications include:

• Protein–Protein Interaction Studies: Use as bait or prey in pull-down assays and co-immunoprecipitation to map the CFAP418 interactome within the ciliary transition zone and connecting cilium.
• Antibody Validation: Serve as a positive control antigen for validating anti-CFAP418 antibodies in Western blotting and ELISA formats.
• Structural and Biophysical Studies: Support circular dichroism, dynamic light scattering, or limited proteolysis experiments to characterize the protein's secondary structure and stability.
• Drug Discovery: Serve as a target protein in small-molecule screening assays aimed at modulating cilia-related pathways implicated in ciliopathies.
• Functional Rescue Experiments: Provide recombinant protein for in vitro complementation assays to study the molecular consequences of disease-associated CFAP418 mutations.

These applications collectively support both mechanistic research into cilia biology and preclinical investigation of therapeutic strategies for CFAP418-associated retinal dystrophies.

Handling, Storage & Quality Considerations

Proper handling and storage of Recombinant Human CFAP418 Protein are essential to preserve protein integrity and experimental reproducibility. The following general guidelines apply; always refer to the product-specific datasheet for confirmed conditions:

• Storage Temperature: Store at −20°C for short-term use or −80°C for long-term storage. Avoid repeated freeze-thaw cycles, which can lead to aggregation and loss of activity.
• Aliquoting: Upon first thaw, divide the protein into single-use aliquots to minimize freeze-thaw events.
• Carrier Protein: Many recombinant protein formulations include BSA or glycerol as a stabilizer; confirm the formulation before designing downstream assays to avoid interference.
• Purity Assessment: Confirm protein purity by SDS-PAGE prior to use in sensitive binding or activity assays. Expected purity for research-grade recombinant proteins is typically ≥85%, but verify with the supplied Certificate of Analysis (CoA).
• Endotoxin Levels: For cell-based assays, verify that endotoxin levels are within acceptable limits as stated on the CoA to prevent confounding inflammatory responses.

Adherence to these practices will ensure data reproducibility and maximize the utility of this recombinant protein across experimental platforms.

Frequently asked questions

What is CFAP418 and why is it important in research?

CFAP418 (Cilia and Flagella Associated Protein 418, also known as C8orf37) is a protein essential for cilia assembly and maintenance in photoreceptor cells and other ciliated tissues. It is important in research because loss-of-function mutations in the CFAP418 gene cause autosomal recessive retinal dystrophies and have been linked to Bardet-Biedl syndrome, making it a key target for understanding ciliopathies and developing potential therapies.

What are the recommended storage conditions for Recombinant Human CFAP418 Protein?

Recombinant Human CFAP418 Protein should generally be stored at −20°C for short-term use or at −80°C for long-term storage. Repeated freeze-thaw cycles should be avoided to prevent protein aggregation and activity loss; it is recommended to aliquot the protein into single-use volumes upon first thaw. Always consult the product-specific datasheet and Certificate of Analysis for confirmed storage conditions and formulation details.

What applications is Recombinant Human CFAP418 Protein suitable for?

Recombinant Human CFAP418 Protein is suitable for a range of research applications including protein-protein interaction studies (pull-down assays, co-immunoprecipitation), antibody validation via Western blot and ELISA, structural and biophysical characterization, small-molecule screening in drug discovery, and in vitro functional rescue experiments. It is particularly relevant for researchers studying cilia biology, ciliopathies, and inherited retinal degenerative diseases.

How should I assess the purity and quality of Recombinant Human CFAP418 Protein before use?

Protein purity can be assessed by running an SDS-PAGE gel and comparing the band pattern to the expected molecular weight. Researchers should review the Certificate of Analysis (CoA) supplied with the product (SKU: rp373155) for confirmed purity levels, endotoxin content, and formulation details. For cell-based assays, verifying endotoxin levels is particularly important to prevent non-specific inflammatory responses that could confound results.

References

1. Estrada-Cuzcano A, et al. Loss-of-function mutations in the gene encoding the cilia- and flagella-associated protein CFAP418 cause Bardet-Biedl syndrome. Nat Genet. 2012.
2. Sharif AS, et al. C8orf37 is required for photoreceptor outer segment disc morphogenesis and outer segment development. Hum Mol Genet. 2018.
3. Roosing S, et al. Mutations in CFAP418 that affect ciliogenesis cause autosomal-recessive retinal dystrophy. Ophthalmology. 2013.

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