Recombinant Human CDCA7L Protein: Production & Usage Protocol

Recombinant Human CDCA7L Protein is a research-grade recombinant protein targeting the cell division cycle associated 7-like (CDCA7L) gene product, supplied by Aladdin Scientific to support studies in cell cycle regulation, transcriptional control, and related signaling pathways.

Product Name

Recombinant Human CDCA7L Protein

SKU

rp375946

Target Protein

CDCA7L (Cell Division Cycle Associated 7-Like)

Species

Human (Homo sapiens)

Application

Cell cycle research, transcription regulation studies, biochemical assays

Storage Recommendation

Avoid repeated freeze-thaw cycles; store in aliquots at -80°C for long-term stability

Background and Biological Significance of CDCA7L

CDCA7L (Cell Division Cycle Associated 7-Like) is a paralog of CDCA7 and belongs to a family of proteins implicated in cell cycle progression and transcriptional regulation. CDCA7L contains a conserved zinc-finger domain that is believed to mediate interactions with chromatin-associated complexes, including components of the HELLS (LSH) chromatin-remodeling pathway.

Research suggests that CDCA7L plays roles in DNA methylation maintenance, genome stability, and the regulation of cell proliferation. Dysregulation of CDCA7L has been associated with immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome-related pathways, making it a protein of significant interest in epigenetics and cancer biology research.

The recombinant form of this protein (SKU: rp375946) enables researchers to study CDCA7L function in vitro, perform binding assays, generate antibodies, and validate experimental tools under controlled conditions.

Reconstitution and Storage Protocol

Reconstitution: Prior to reconstitution, briefly centrifuge the vial to collect lyophilized material at the bottom. Reconstitute the protein in sterile, deionized water or an appropriate buffer (e.g., PBS, pH 7.4) to a working concentration as recommended by the product datasheet. Gently pipette to mix; do not vortex vigorously, as this may denature the protein.

Working Concentration: Prepare working aliquots at the concentration required for your specific assay. Consult the product datasheet (referenced as absea#PP-9580) for lot-specific concentration and purity information.

Short-term Storage: Reconstituted protein may be stored at 4°C for up to 48–72 hours if used within that timeframe.

Long-term Storage: For long-term storage, aliquot the reconstituted protein to minimize freeze-thaw cycles and store at -80°C. Adding a carrier protein such as 0.1% BSA may improve stability during long-term storage.

Important: Avoid repeated freeze-thaw cycles, as this may significantly reduce protein activity and integrity.

Recommended Experimental Applications

Recombinant Human CDCA7L Protein (rp375946) is suitable for a range of in vitro research applications, including but not limited to:

• SDS-PAGE and Western Blot: Use as a positive control to confirm antibody specificity or to validate detection systems targeting CDCA7L.
• ELISA Development: Serve as a standard or capture antigen in sandwich or competitive ELISA formats for quantifying CDCA7L in biological samples.
• Protein–Protein Interaction Studies: Employ in co-immunoprecipitation or pull-down assays to identify binding partners of CDCA7L within chromatin-remodeling or cell cycle complexes.
• Immunogen for Antibody Production: Use as an immunogen to generate monoclonal or polyclonal antibodies against human CDCA7L.
• Biochemical and Functional Assays: Incorporate into assays examining CDCA7L's role in DNA methylation, chromatin remodeling, or transcriptional regulation.

Always titrate the protein for each specific application to determine optimal working conditions.

Quality Control and Handling Precautions

Purity Verification: Each lot of Recombinant Human CDCA7L Protein undergoes quality control assessment. Refer to the Certificate of Analysis (CoA) provided with each shipment for lot-specific purity, concentration, and endotoxin data.

Endotoxin: For cell-based assays, confirm the endotoxin level meets your experimental requirements. Consult the CoA for lot-specific endotoxin values.

Handling: Always handle the protein on ice to minimize degradation. Use low-protein-binding tubes and pipette tips to reduce adsorption losses.

Personal Protective Equipment: As with all research reagents, wear appropriate PPE including gloves and laboratory coat. Although CDCA7L recombinant protein is not classified as hazardous, standard laboratory safety practices apply.

Contamination Prevention: Use aseptic technique throughout reconstitution and aliquoting to prevent microbial contamination, which can degrade protein integrity and compromise experimental results.

Troubleshooting Common Issues

No Signal in Western Blot: Confirm that the protein was loaded correctly and that the primary antibody recognizes the recombinant form. Check the CoA for molecular weight and verify gel percentage is appropriate for the expected protein size.

Low Activity in Functional Assays: Excessive freeze-thaw cycles are a common cause of reduced activity. Ensure aliquots were prepared during the initial reconstitution. Verify the assay buffer composition is compatible with CDCA7L stability.

Protein Aggregation: If precipitation is observed, centrifuge briefly (10,000 × g, 5 min, 4°C) to remove aggregates before use. Consider adding a mild detergent or adjusting salt concentration to improve solubility.

High Background in ELISA: Optimize blocking conditions and antibody dilutions. Ensure the recombinant protein stock is free from aggregates that could cause non-specific binding.

If issues persist, contact Aladdin Scientific technical support with your lot number and experimental conditions for further assistance.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human CDCA7L Protein?

Recombinant Human CDCA7L Protein is generally reconstituted in sterile deionized water or PBS (pH 7.4). Always refer to the lot-specific Certificate of Analysis and product datasheet (absea#PP-9580) for the manufacturer's recommended reconstitution conditions, as buffer requirements may vary by formulation.

How should Recombinant Human CDCA7L Protein be stored to maintain activity?

For short-term use, store at 4°C for up to 48–72 hours. For long-term storage, aliquot the reconstituted protein and store at -80°C to avoid repeated freeze-thaw cycles. Adding a carrier protein such as 0.1% BSA may help maintain stability.

What applications is Recombinant Human CDCA7L Protein (rp375946) validated for?

This recombinant protein is suitable for applications including SDS-PAGE, Western blot positive control, ELISA standard or antigen, protein-protein interaction studies, and use as an immunogen for antibody generation. Researchers should validate the protein for each specific application and consult the product datasheet for confirmed activities.

What is the biological function of CDCA7L?

CDCA7L is a paralog of CDCA7 involved in cell cycle regulation, transcriptional control, and chromatin remodeling, particularly through interactions with the HELLS/LSH complex. It is implicated in DNA methylation maintenance and genome stability, and has been linked to ICF syndrome-related pathways.

How can I troubleshoot low signal when using CDCA7L protein in a Western blot?

First, verify that the protein was loaded at an appropriate quantity and that the antibody recognizes the recombinant form. Check the molecular weight on the CoA and ensure the gel percentage is suitable. Also confirm the protein has not undergone excessive freeze-thaw cycles, which can reduce antigenicity.

References

1. Absea Biotechnology. Product Data Sheet: Recombinant Human CDCA7L Protein, Reference PP-9580.
2. Jenness C, et al. HELLS and CDCA7 comprise a bipartite nucleosome remodeling complex defective in ICF syndrome. Proceedings of the National Academy of Sciences. 2018.
3. Thijssen PE, et al. Mutations in CDCA7 and HELLS cause immunodeficiency-centromeric instability-facial anomalies syndrome. Nature Genetics. 2015.

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