Recombinant Human CFAP57 Protein: Handling & Usage Protocol

Recombinant Human CFAP57 Protein is a research-grade reagent designed to support investigations into cilia-associated protein function, motile cilia assembly, and related cellular processes.

Product Name

Recombinant Human CFAP57 Protein

SKU

rp375991

Protein Origin

Human (Homo sapiens)

Protein Type

Recombinant, cilia and flagella associated protein

Recommended Storage

Store lyophilized protein at -20°C or -80°C; avoid repeated freeze-thaw cycles

Intended Use

Research use only (RUO); not for diagnostic or therapeutic use

Background: CFAP57 and Its Biological Role

CFAP57 (Cilia and Flagella Associated Protein 57) is a protein encoded by the CFAP57 gene in humans. As a member of the cilia and flagella associated protein family, CFAP57 is believed to play a structural or regulatory role in the assembly and function of motile cilia and flagella. Motile cilia are essential organelles found on epithelial surfaces of the respiratory tract, brain ventricles, and reproductive organs, where they drive fluid movement and cellular transport.

Dysfunction or absence of proteins in the CFAP family has been associated with primary ciliary dyskinesia (PCD) and related ciliopathies, conditions characterized by impaired cilia motility leading to chronic respiratory infections, laterality defects, and infertility. Research into CFAP57 and its interaction partners contributes to the broader understanding of cilia biology and the molecular basis of ciliopathies.

Reconstitution Protocol

Follow these steps to reconstitute Recombinant Human CFAP57 Protein (rp375991) for downstream experiments:

1. Equilibration: Allow the vial to equilibrate to room temperature for 5–10 minutes before opening to minimize condensation and protein aggregation.
2. Reconstitution buffer: Reconstitute the lyophilized protein in sterile deionized water or an appropriate buffer (e.g., PBS, pH 7.4) as specified on the product datasheet. Gently swirl or pipette to mix — do not vortex vigorously.
3. Concentration: Reconstitute to the concentration recommended on the Certificate of Analysis (CoA). Refer to the product datasheet for the specific working concentration range.
4. Centrifugation: Briefly centrifuge the vial (1,000 × g, 30 seconds) before opening to ensure all lyophilized material is at the bottom of the vial.
5. Aliquoting: Prepare single-use aliquots immediately after reconstitution to avoid repeated freeze-thaw cycles, which can compromise protein activity and integrity.

Note: Specific reconstitution volumes and buffer recommendations are provided on the product datasheet and Certificate of Analysis supplied with each lot. Always refer to lot-specific documentation.

Storage and Stability Guidelines

Proper storage is critical to maintaining the activity and integrity of Recombinant Human CFAP57 Protein:

• Lyophilized form: Store at -20°C or -80°C in a frost-free freezer. Lyophilized protein is generally stable for up to 12 months from the date of receipt when stored correctly.
• Reconstituted protein: After reconstitution, store working aliquots at -80°C. For short-term use (within 1 week), storage at 4°C may be acceptable, but activity should be verified.
• Avoid freeze-thaw cycles: Each freeze-thaw cycle can degrade protein structure and reduce biological activity. Single-use aliquots are strongly recommended.
• Carrier protein: If the protein is provided without a carrier, consider adding bovine serum albumin (BSA) to a final concentration of 0.1% to improve stability during storage, unless incompatible with your assay.

Always consult the product-specific datasheet and CoA for lot-specific stability data and shelf life information.

Experimental Applications

Recombinant Human CFAP57 Protein is suitable for a variety of research applications in cilia biology and related disciplines. Typical uses include:

• Protein–protein interaction studies: Use as bait or prey in pull-down assays, co-immunoprecipitation (co-IP), or surface plasmon resonance (SPR) experiments to identify CFAP57 binding partners.
• Antibody production and validation: Serve as an immunogen for generating anti-CFAP57 antibodies, or as a positive control antigen in ELISA and Western blot validation experiments.
• Functional/biochemical assays: Investigate the biochemical properties of CFAP57, including its role in axonemal assembly or interaction with dynein regulatory complexes.
• Structural studies: Suitable as a substrate for structural biology approaches such as cryo-electron microscopy (cryo-EM) or X-ray crystallography (subject to purity and buffer compatibility).
• Cell-based assays: Use in cell-free or reconstituted systems to study cilia assembly dynamics.

Note: Optimal working concentrations and assay conditions should be determined empirically for each specific application. This product is for research use only and is not intended for clinical or diagnostic applications.

Quality Control and Troubleshooting

Each lot of Recombinant Human CFAP57 Protein undergoes quality control testing prior to release. Typical QC assessments may include SDS-PAGE for purity evaluation and Western blot for identity confirmation. Refer to the CoA provided with your specific lot for detailed QC data.

Common Troubleshooting Tips

• Low protein activity: Verify that the protein has not undergone multiple freeze-thaw cycles. Check reconstitution buffer composition and pH. Ensure incubation conditions (temperature, time) are optimized for your assay.
• Protein precipitation: If precipitation is observed after reconstitution, gently centrifuge (10,000 × g, 5 minutes at 4°C) and use the supernatant. Consider adding a compatible carrier protein or detergent at low concentration if aggregation persists.
• Unexpected bands on SDS-PAGE or Western blot: Minor bands may represent post-translational modifications, degradation products, or host-cell-derived impurities within acceptable QC limits. Contact Aladdin Scientific technical support if results are inconsistent with the CoA.
• No signal in functional assay: Confirm that the assay format and detection method are validated for CFAP57. Optimize protein concentration and assay buffer conditions.

For further technical assistance, please contact Aladdin Scientific customer support.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human CFAP57 Protein?

The recommended reconstitution buffer is typically sterile deionized water or PBS (pH 7.4), as specified on the product datasheet and Certificate of Analysis supplied with each lot. Always refer to the lot-specific documentation for precise instructions, as buffer recommendations may vary by formulation.

How should I store Recombinant Human CFAP57 Protein to maintain its activity?

Store the lyophilized protein at -20°C or -80°C in a frost-free freezer. After reconstitution, prepare single-use aliquots and store at -80°C to avoid repeated freeze-thaw cycles, which can significantly reduce protein activity and structural integrity. Short-term storage at 4°C (up to one week) may be acceptable but should be validated for your specific application.

What applications is Recombinant Human CFAP57 Protein validated for?

Recombinant Human CFAP57 Protein is suitable for research applications including protein–protein interaction studies (e.g., pull-down, co-IP, SPR), antibody production and validation (ELISA, Western blot), functional and biochemical assays, and structural biology studies. Specific application validations and recommended concentrations are detailed in the product datasheet. This product is for research use only.

Is Recombinant Human CFAP57 Protein suitable for clinical or diagnostic use?

No. Recombinant Human CFAP57 Protein (rp375991) is strictly for research use only (RUO) and is not approved or intended for use in clinical diagnosis, therapeutic applications, or any procedures involving humans or animals in a clinical setting.

What should I do if I observe protein precipitation after reconstitution?

If precipitation is observed, gently centrifuge the reconstituted solution at 10,000 × g for 5 minutes at 4°C and use the clarified supernatant. If aggregation persists, consider adding a compatible carrier protein such as 0.1% BSA or a low concentration of a non-ionic detergent. Contact Aladdin Scientific technical support if the issue continues.

References

1. Horani A, Ferkol TW. Understanding Primary Ciliary Dyskinesia and Other Ciliopathies. J Pediatr. 2021;230:15-22.e1. doi:10.1016/j.jpeds.2020.11.040
2. Reiter JF, Leroux MR. Genes and molecular pathways underpinning ciliopathies. Nat Rev Mol Cell Biol. 2017;18(9):533-547. doi:10.1038/nrm.2017.60
3. Wallmeier J, et al. Motile ciliopathies. Nat Rev Dis Primers. 2020;6(1):77. doi:10.1038/s41572-020-0209-6

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