Background: CDCA3 Biology and Research Relevance

CDCA3, also known as Cell Division Cycle Associated 3 (also referred to as Tome-1 in some literature), is a cell cycle regulatory protein that plays an important role in controlling the G2/M transition. It functions as an adaptor subunit within the SCF (Skp1–Cullin–F-box) E3 ubiquitin ligase complex, where it facilitates the ubiquitin-mediated degradation of key substrates such as Wee1 kinase, thereby promoting mitotic entry.

CDCA3 expression has been reported to be elevated in multiple cancer types, making it a subject of interest in oncology research and a potential biomarker or therapeutic target. Research scientists studying mitotic checkpoint mechanisms, cell proliferation, or cancer biology frequently require high-quality recombinant CDCA3 protein for in vitro biochemical experiments, structural studies, and assay development.

Recombinant Human CDCA3 Protein from Aladdin Scientific (SKU: rp374385) provides researchers with a reliable, ready-to-use reagent for these research applications.

Reconstitution and Handling Protocol

Proper handling of recombinant proteins is essential to maintain biological activity and ensure reproducible experimental results. Follow the steps below when working with Recombinant Human CDCA3 Protein:

1. Centrifugation before opening: Briefly centrifuge the vial (e.g., 5–10 seconds at low speed) to collect any lyophilized or frozen material at the bottom before opening.
2. Reconstitution: Reconstitute the protein in an appropriate buffer (typically sterile deionized water or PBS, pH 7.4, supplemented with a carrier protein such as 0.1% BSA if required for stability). Refer to the product-specific datasheet for the recommended reconstitution volume and buffer composition.
3. Gentle mixing: Mix gently by pipetting or slow vortexing. Do not vigorously vortex, as this may denature the protein.
4. Aliquoting: Prepare single-use aliquots immediately after reconstitution to avoid repeated freeze-thaw cycles, which can degrade protein integrity and activity.
5. Storage after reconstitution: Store short-term aliquots at 4°C for up to 1 week, or at -20°C to -80°C for longer-term storage. Consult the datasheet for product-specific stability data.

Note: Specific reconstitution conditions (buffer, volume, additives) should always be confirmed in the product datasheet supplied with the reagent. The above represents general best-practice guidance for recombinant proteins.

Recommended Applications and Experimental Guidelines

Recombinant Human CDCA3 Protein is suitable for a range of in vitro research applications. The following guidelines outline common uses and considerations for each:

• Biochemical Assays (e.g., ubiquitination assays): CDCA3 can be used as a component or substrate in reconstituted SCF complex assays to study its role in ubiquitin-mediated proteolysis. Optimize protein concentrations empirically for each assay format.
• Protein–Protein Interaction Studies (co-IP, pull-down, SPR): Use CDCA3 protein as bait or prey to identify binding partners involved in cell cycle regulation. Ensure appropriate controls, including negative controls with unrelated proteins, are included.
• Western Blot Standard: Recombinant CDCA3 protein serves as a molecular weight and immunoreactivity standard for anti-CDCA3 antibody validation in Western blot experiments.
• ELISA Development: Can be used as a coating or detection antigen for sandwich ELISA development targeting CDCA3.
• Cell-Free Expression and Functional Studies: Suitable for in vitro functional assays where addition of exogenous recombinant protein is required to complement or reconstitute activity in cell-free systems.

Always titrate the protein and optimize conditions for each specific assay. Refer to published literature on CDCA3 biology for assay design guidance.

Quality, Safety, and Regulatory Considerations

When working with Recombinant Human CDCA3 Protein, researchers should be aware of the following quality and safety considerations:

• Purity and Quality: The recombinant protein is manufactured under controlled conditions. Review the Certificate of Analysis (CoA) provided with each lot for purity grade, endotoxin level, and activity data specific to that lot.
• Endotoxin Content: If the protein is to be used in cell-based assays or in vivo studies, verify that endotoxin levels are within acceptable limits for the intended application. Consult the product datasheet or contact Aladdin Scientific technical support for lot-specific endotoxin data.
• Biosafety: Recombinant Human CDCA3 Protein is derived from human gene sequences expressed in a recombinant host. Handle according to your institution's biosafety guidelines for recombinant proteins. Consult the Safety Data Sheet (SDS) provided by Aladdin Scientific before use.
• Research Use Only: This product is intended for in vitro research use only. It is not approved for diagnostic, therapeutic, or veterinary applications.
• Lot-to-Lot Consistency: For studies requiring strict reproducibility, purchase sufficient quantity from a single lot where possible. Contact Aladdin Scientific for lot reservation inquiries.