Recombinant Human Cyr61

Recombinant human CYR61 (also known as CCN1) is a valuable research tool with diverse applications across multiple biological systems. Here are the key reasons to incorporate it into your research:

Cellular Adhesion and Migration

Recombinant CYR61 promotes cell adhesion and stimulates cell migration in fibroblasts and endothelial cells. The protein interacts with multiple integrin receptors on cell surfaces—including αvβ3, αvβ5, α6β1, and αIIbβ3—triggering intracellular signaling pathways that enhance cell motility and proliferation. This makes it particularly useful for studying cell-cell interactions and migration mechanisms in various tissue contexts.

Angiogenesis Research

CYR61 is a potent angiogenic factor that significantly enhances endothelial cell proliferation, migration, and tube formation. In vitro studies demonstrate that CYR61 treatment increases tubular network formation and vessel maturation, facilitating the recruitment and assembly of endothelial cells into neovessel structures. This capability makes it an excellent tool for investigating angiogenic mechanisms and vascular development.

Growth Factor Signaling Studies

Recombinant CYR61 augments growth factor-induced DNA synthesis in fibroblasts and endothelial cells, allowing you to study the crosstalk between CYR61 signaling and other growth factor pathways. This is particularly valuable for understanding how matricellular proteins modulate mitogenic responses.

Bone Regeneration and Fracture Repair

CYR61 delivery accelerates neovascularization during bone fracture repair without altering cartilage or bone formation directly. This selective enhancement of angiogenesis makes it useful for studying the angiogenic-osteogenic crosstalk and for developing therapeutic strategies for fractures with limited blood vessel supply.

Cancer Research and Biomarker Development

CYR61 exhibits context-dependent roles in cancer, acting as both a tumor promoter and suppressor depending on cancer type. Its involvement in extracellular matrix remodeling, immune modulation, and integrin-mediated signaling makes it valuable for investigating cancer progression mechanisms and exploring its potential as a diagnostic biomarker. Additionally, CYR61 enhances the efficacy of CAR-T cell approaches targeting αvβ3 integrin, improving their cytotoxic efficacy against tumor cells.

Immunological Applications

Recent research highlights CYR61's role in regulating cytokine and chemokine production through autocrine and paracrine feedback mechanisms, suggesting its utility in studying immune surveillance and inflammatory processes.

Technical Advantages

Recombinant CYR61 is typically produced as a 40-45 kDa matricellular glycoprotein with high sequence conservation across species (93% identity between human, mouse, and rat forms). The protein can be used in multiple experimental formats including bioassays, cell-based assays, and in vivo delivery studies, providing flexibility for various research applications.

Yes, recombinant human Cyr61 (CCN1) can be used as a standard for quantification or calibration in ELISA assays, provided that the recombinant protein is suitable for your specific assay format and detection system.

Key Considerations:

• Purity and Form: Ensure the recombinant Cyr61 is highly purified and in a form compatible with your ELISA (e.g., carrier-free if possible, to avoid interference from additives like BSA).
• Concentration and Dilution: Prepare a dilution series of the recombinant protein to generate a standard curve. This curve will allow you to interpolate the concentration of Cyr61 in your samples.
• Compatibility: The recombinant protein should be recognized by the capture and detection antibodies used in your ELISA kit. Most commercial ELISA kits are validated for use with their own standards, but recombinant proteins from reputable suppliers (such as R&D Systems, Novus Biologicals, or Abcam) are generally suitable.
• Validation: It is good practice to validate the recombinant protein as a standard by comparing its performance to the kit’s provided standard, if available.

Suppliers Offering Recombinant Human Cyr61:

• R&D Systems: Recombinant Human Cyr61/CCN1 Fc Chimera Protein (carrier-free or with BSA options).
• Novus Biologicals: Recombinant Human Cyr61/CCN1 Protein (E. coli expressed, non-glycosylated).
• Abcam: Recombinant Human CYR61 protein (various forms available).

Summary:

Yes, recombinant human Cyr61 can be used as a standard in ELISA assays for quantification, as long as it is compatible with your assay system and properly validated. Always check the product datasheet for recommended usage and compatibility.

Recombinant Human Cyr61 (CCN1) has been validated in published research for applications including angiogenesis assays, cell migration and adhesion studies, tumorigenesis models, wound and bone repair, immune modulation, and as a biomarker in cancer prognosis.

Key validated applications include:

• Angiogenesis assays: Cyr61 stimulates endothelial cell proliferation, migration, and tube formation in vitro, and induces neovascularization in vivo (e.g., rat cornea model, bone fracture repair).
• Cell migration and adhesion: Cyr61 promotes directed migration of endothelial and other cell types, often via integrin-dependent pathways (notably αvβ3 and αvβ5).
• Tumorigenesis and cancer progression: Cyr61 is used in models to study tumor growth, vascularization, and metastasis, and its expression correlates with tumor progression in multiple cancer types (breast, renal, gastric, colorectal, glioma, lung).
• Wound healing and tissue repair: Cyr61 is upregulated in injured skin and bone, where it induces growth factors and cytokines involved in repair; recombinant Cyr61 accelerates neovascularization during bone fracture repair.
• Immune modulation: Cyr61 regulates cytokine and chemokine production, modulates immune cell migration, and participates in immune surveillance, including vascular inflammation models.
• Bioassays and flow cytometry: Used to assess cellular responses such as adhesion, migration, and immune cell patrolling in vitro and ex vivo.
• Biomarker and therapeutic target: Cyr61 is explored as a diagnostic biomarker for cancer prognosis and as a target for therapies, including CAR-T cell approaches.

Experimental protocols and models:

• In vitro: Endothelial tube formation, cell migration/adhesion assays, chondrogenesis and osteogenesis differentiation assays, cytokine induction studies.
• In vivo: Tumor xenograft models, bone fracture repair models, corneal neovascularization assays.
• Ex vivo: Flow cytometry of immune cell populations, organoid cultures.

Summary of validated research uses:

• Angiogenesis and vascular biology
• Cancer biology and metastasis
• Cell migration, adhesion, and signaling
• Wound and bone healing
• Immune cell modulation and inflammation
• Biomarker development and therapeutic targeting

These applications are supported by multiple peer-reviewed studies and reviews, demonstrating the versatility of recombinant human Cyr61 in both basic and translational research contexts.

To properly reconstitute and prepare Recombinant Human Cyr61 (CCN1) protein for cell culture experiments, follow these general best practices based on manufacturer guidelines and scientific protocols:

Reconstitution

1. Centrifuge the vial: Before opening, briefly centrifuge the lyophilized protein vial to ensure all powder is at the bottom.
2. Reconstitution solvent: Reconstitute the protein in sterile distilled water or a buffer as specified by the manufacturer (e.g., PBS, pH 7.4). For most Cyr61 proteins, sterile distilled water is recommended.
3. Concentration: Reconstitute to a concentration between 0.1–0.5 mg/mL (or as specified by the product datasheet). For example, if the vial contains 10 µg of protein, add 20–100 µL of solvent.
4. Gentle mixing: Gently swirl or pipette up and down to dissolve the protein. Do not vortex or shake vigorously to avoid denaturation.
5. Incubation: Allow the vial to sit at room temperature for 15–30 minutes with gentle agitation to ensure complete dissolution.

Preparation for Cell Culture

1. Further dilution: Dilute the reconstituted protein to the desired working concentration in cell culture medium or an appropriate buffer.
2. Carrier protein (optional): For long-term storage or to stabilize the protein, consider adding a carrier protein (e.g., 0.1% BSA, 5% HSA, or 5% trehalose) to the reconstituted solution.
3. Aliquoting: Aliquot the reconstituted protein into small volumes to minimize freeze-thaw cycles.
4. Storage: Store aliquots at –20°C to –80°C for long-term use. After reconstitution, the protein is typically stable for 3 months at –20°C and 1 week at 2–8°C.

Application in Cell Culture

• Immobilization (if required): For adhesion or migration assays, coat plates with Cyr61 at a concentration of 1.5 µg/mL in PBS, incubate at 37°C for 1–2 hours, then wash before adding cells.
• Direct addition: For proliferation or signaling studies, add the diluted Cyr61 directly to the culture medium at the desired concentration.

Important Notes

• Avoid repeated freeze-thaw cycles to maintain protein activity.
• Sterile technique: Always use sterile solutions and equipment to prevent contamination.
• Check the datasheet: Always refer to the specific product datasheet for exact reconstitution and storage instructions, as protocols may vary slightly between manufacturers.

By following these steps, you can ensure the proper reconstitution and preparation of Recombinant Human Cyr61 protein for reliable and reproducible cell culture experiments.