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.