Recombinant Human CDC5 Protein: Research Guide & Applications

Recombinant Human CDC5 Protein is a key research reagent enabling scientists to investigate the molecular mechanisms of pre-mRNA splicing, cell cycle regulation, and DNA damage checkpoint responses in human cells.

Product Name

Recombinant Human CDC5 Protein

SKU

rp375930

Also Known As

CDC5L, PCDC5RP, Cdc5-like protein, Cell division cycle 5-like protein

Biological Function

Component of the Prp19 splicing complex; role in pre-mRNA splicing and DNA damage checkpoint activation

Species Origin

Human (Homo sapiens)

Application Areas

Cell cycle research, RNA splicing studies, DNA damage response assays, protein interaction studies

Supplier Reference

absea#PP-9565

What Is CDC5 and Why Does It Matter?

CDC5 (Cell Division Cycle 5-like protein, also designated CDC5L) is a conserved nuclear protein that plays dual and interconnected roles in eukaryotic cells. It is an essential component of the Prp19 complex (also called the NineTeen Complex, NTC), a multi-subunit assembly required for the activation and catalytic steps of pre-mRNA splicing. Beyond RNA processing, CDC5L has been identified as a critical mediator of the DNA damage checkpoint response, functioning in the signaling cascade that arrests cell cycle progression in response to genotoxic stress.

The protein contains a Myb-like DNA-binding domain and coiled-coil regions that facilitate protein–protein interactions within the spliceosome. Mutations or dysregulation of CDC5L have been associated with genomic instability, making it a relevant target in cancer biology research. Recombinant Human CDC5 Protein produced under controlled conditions enables reproducible in vitro investigations of these pathways.

Recombinant Human CDC5 Protein: Key Characteristics and Quality Considerations

Recombinant Human CDC5 Protein (Aladdin Scientific SKU: rp375930; supplier reference absea#PP-9565) is manufactured using recombinant expression systems to provide researchers with a well-characterized reagent for functional and biochemical studies. When evaluating recombinant proteins, researchers should confirm the following quality attributes from the certificate of analysis (CoA):

• Purity: Assessed by SDS-PAGE and/or HPLC; high purity minimizes non-specific effects in downstream assays.
• Identity: Confirmed by Western blot or mass spectrometry using anti-CDC5L antibodies or peptide mapping.
• Biological Activity: Where applicable, functional assays (e.g., protein–protein interaction assays with known CDC5L binding partners such as PLRG1 or BCAS2) confirm activity.
• Endotoxin Level: Low endotoxin content is essential for cell-based assays to avoid confounding inflammatory responses.

Always request and review the CoA for each lot before use. Contact Aladdin Scientific technical support for lot-specific data sheets.

Recommended Handling, Storage, and Reconstitution Protocol

Proper handling of recombinant proteins is essential to preserve activity and ensure experimental reproducibility. The following general guidelines apply to Recombinant Human CDC5 Protein; always defer to the product-specific CoA for definitive instructions:

1. Receipt and Inspection: Upon receipt, inspect the vial for integrity. Record the lot number and expiry date. If the product is shipped on dry ice, allow it to equilibrate briefly at the recommended storage temperature before opening.
2. Storage: Store lyophilized protein at –20 °C or –80 °C as specified on the CoA. Avoid repeated freeze–thaw cycles, which can cause aggregation and loss of activity. Aliquot reconstituted protein into single-use volumes.
3. Reconstitution: Reconstitute lyophilized protein in the volume and buffer specified on the CoA (typically sterile water or a compatible buffer such as PBS with a carrier protein). Allow the vial to equilibrate to room temperature, add reconstitution buffer dropwise, and gently swirl — do not vortex.
4. Working Concentrations: Determine optimal working concentrations empirically for each assay type. Titration experiments are recommended when using the protein for the first time.
5. Protein Stability: Addition of a carrier protein (e.g., 0.1% BSA) to the reconstitution or dilution buffer can enhance stability during storage and assay incubations.

Note: Specific storage conditions, recommended buffers, and concentration data are provided on the lot-specific CoA. Contact Aladdin Scientific for the most current product documentation.

Research Applications of Recombinant Human CDC5 Protein

Recombinant Human CDC5 Protein is a versatile reagent applicable across several research domains:

• Pre-mRNA Splicing Studies: CDC5L is a structural and functional component of the Prp19/NTC complex. The recombinant protein can be used in in vitro splicing reconstitution experiments, pull-down assays to map interactions with other spliceosomal components, and competition assays to probe spliceosome assembly.
• Cell Cycle and DNA Damage Checkpoint Research: CDC5L participates in checkpoint signaling following DNA double-strand breaks. Recombinant CDC5 protein can serve as a substrate or binding partner in kinase assays (e.g., with ATR/ATM pathway components) and in co-immunoprecipitation validation experiments.
• Protein–Protein Interaction (PPI) Studies: Surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and ELISA-based binding assays can use purified CDC5 protein to quantify interactions with binding partners such as PLRG1, BCAS2, and CDC5L-associated factors.
• Antibody Validation and Immunoassay Development: Recombinant CDC5 protein serves as a positive control antigen for Western blot, ELISA, and immunofluorescence antibody validation.
• Drug Discovery and Target Validation: Given the link between CDC5L dysregulation and cancer, the recombinant protein supports compound screening assays aimed at disrupting spliceosome assembly or CDC5L-dependent checkpoint signaling.

Troubleshooting Common Issues with Recombinant CDC5 Protein Assays

Researchers may encounter the following issues when working with Recombinant Human CDC5 Protein. The guidance below reflects general best practices for recombinant protein reagents:

Issue	Likely Cause	Recommended Action
Protein aggregation or precipitation	Freeze–thaw cycling; incorrect reconstitution buffer; concentration too high	Aliquot into single-use volumes; use carrier protein (0.1% BSA); centrifuge at 10,000 × g for 5 min before use and use supernatant
Low or absent signal in binding assay	Protein denaturation; incompatible buffer conditions; incorrect assay format	Confirm protein integrity by SDS-PAGE; optimize pH, salt, and detergent conditions; include positive control
High background in ELISA	Non-specific binding; insufficient blocking	Optimize blocking buffer (BSA vs. non-fat milk); increase wash steps; titrate protein concentration
Inconsistent results between lots	Normal lot-to-lot biological variability	Request lot-specific CoA; aliquot and store all lots identically; include internal reference standard

For persistent technical issues, contact Aladdin Scientific technical support with your lot number, assay description, and CoA data.

Frequently asked questions

What is the biological function of CDC5 (CDC5L) protein in human cells?

CDC5L is a multifunctional nuclear protein with two primary roles: it is an integral component of the Prp19/NTC spliceosomal complex required for pre-mRNA splicing, and it participates in the DNA damage checkpoint response, helping to coordinate cell cycle arrest following genotoxic stress. Its Myb-like domain facilitates DNA binding and protein–protein interactions within these complexes.

How should I store Recombinant Human CDC5 Protein to maintain its activity?

Lyophilized Recombinant Human CDC5 Protein should be stored at –20 °C or –80 °C as specified on the lot-specific certificate of analysis. After reconstitution, divide the protein into single-use aliquots to avoid repeated freeze–thaw cycles, which cause aggregation and activity loss. Adding a carrier protein such as 0.1% BSA to the storage buffer can further stabilize the protein.

What assay formats are compatible with Recombinant Human CDC5 Protein?

Recombinant Human CDC5 Protein is compatible with multiple assay formats including Western blot (as a positive control antigen), ELISA-based binding assays, pull-down and co-immunoprecipitation experiments, surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and in vitro kinase assays. Optimal working concentrations should be determined empirically for each assay format.

Is Recombinant Human CDC5 Protein suitable for cell-based assays?

Recombinant Human CDC5 Protein can be used in cell-based contexts, but low endotoxin content is critical to avoid confounding innate immune or inflammatory responses. Always verify the endotoxin specification on the CoA before use in cell culture assays. For intracellular applications, a suitable protein delivery strategy (e.g., transfection reagent or electroporation) may be required.

How do I troubleshoot low binding signal when using CDC5 protein in a pull-down assay?

Low binding signal in pull-down assays typically results from protein denaturation, incompatible buffer conditions, or suboptimal protein concentration. First, confirm protein integrity by running an SDS-PAGE gel. Then, optimize buffer pH, salt concentration, and detergent type to match the native interaction conditions of CDC5L. Include a known positive control (e.g., a validated CDC5L binding partner) to confirm assay functionality before troubleshooting the target interaction.

References

1. Ajuh P, et al. Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry. EMBO J. 2000;19(23):6569-6581. doi:10.1093/emboj/19.23.6569
2. Zhang N, et al. Identification of a new human cannabinoid receptor. J Neurochemistry. — [Placeholder: original CDC5L checkpoint reference needed]
3. Mu R, et al. Depletion of pre-mRNA splicing factor Cdc5L inhibits mitotic progression and triggers mitotic catastrophe. Cell Death Dis. 2014.
4. Aladdin Scientific. Recombinant Human CDC5 Protein, SKU rp375930, Product Datasheet. Aladdin Scientific.

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