Recombinant Human WISP-1

Recombinant Human WISP-1 (CCN4) is used in research applications to investigate its roles in cell proliferation, differentiation, angiogenesis, tissue regeneration, and cancer biology. Its recombinant form allows for controlled, reproducible studies of WISP-1’s biological functions in vitro and in vivo.

Key scientific applications and rationale:

• Mesenchymal Differentiation: WISP-1 promotes proliferation and lineage-specific differentiation of mesenchymal progenitors, including osteoblasts and chondrocytes, making it valuable for studies on bone and cartilage development, repair, and tissue engineering.
• Angiogenesis: WISP-1 acts as a pro-angiogenic factor, enhancing VEGF-A expression and endothelial cell survival. It is implicated in neovascularization in both physiological (e.g., myocardial infarction recovery) and pathological (e.g., tumor growth) contexts.
• Cancer Research: WISP-1 regulates cell migration, invasion, and metastasis in various cancers by modulating signaling pathways such as integrin αvβ3/FAK/c-Src/EGFR/ERK/HIF1-α. It is used to study tumor microenvironment interactions and angiogenic mechanisms.
• Apoptosis and Cell Survival: WISP-1 attenuates p53-mediated apoptosis and upregulates anti-apoptotic proteins like Bcl-X(L) via AKT activation, relevant for research on cell survival under stress or DNA damage.
• Cardiac and Vascular Biology: WISP-1 is upregulated after myocardial infarction and regulates cardiac endothelial signaling, supporting its use in cardiovascular research.
• Bone Tissue Engineering: WISP-1 signaling drives bone formation and may be harnessed for cell therapy and bone defect healing studies.

Experimental uses:

• Cell culture assays: To study effects on proliferation, differentiation, migration, and survival of various cell types (e.g., mesenchymal stem cells, osteoblasts, endothelial cells).
• Bioassays: For quantifying WISP-1 activity or its downstream effects (e.g., VEGF-A induction, angiogenesis).
• Disease modeling: In vitro and in vivo models of bone regeneration, cancer progression, and cardiovascular injury.

Best practices:

• Use carrier-free recombinant WISP-1 for applications sensitive to additives (e.g., signaling studies, ELISA standards).
• Confirm protein purity and activity via SDS-PAGE, HPLC, and functional assays.
• Select appropriate concentrations and exposure times based on published protocols and cell type sensitivity.

Summary:
Recombinant Human WISP-1 is a versatile tool for dissecting molecular mechanisms in tissue development, regeneration, angiogenesis, and cancer, enabling precise manipulation of WISP-1 signaling in controlled experimental systems.

Yes, recombinant Human WISP-1 protein can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay's antibodies and format.

Most commercial Human WISP-1 ELISA kits use recombinant Human WISP-1 as the reference standard to generate the standard curve for quantification. The standard is typically supplied as a lyophilized or liquid recombinant protein, which is reconstituted and serially diluted to create a calibration curve. The concentration of WISP-1 in unknown samples is then interpolated from this curve.

Key considerations for use:

• Ensure the recombinant WISP-1 standard matches the isoform and sequence recognized by your ELISA antibodies, as WISP-1 has multiple splice variants.
• Use the recommended diluent (often provided with the kit) for standard preparation to avoid matrix effects.
• Store the recombinant standard at -20°C or -80°C as instructed to maintain stability.
• Confirm that your ELISA kit is validated for both natural and recombinant WISP-1, which is standard for most commercial kits.

Best practices:

• Prepare a fresh standard curve for each assay run.
• Avoid repeated freeze-thaw cycles of the recombinant standard to prevent degradation.
• Use the standard curve to interpolate sample concentrations, ensuring all samples and standards are treated identically.

In summary, recombinant Human WISP-1 is widely accepted and routinely used as a calibration standard in ELISA quantification, provided it is compatible with your assay system and handled according to best practices.

Recombinant Human WISP-1 has been validated in published research for several applications, primarily in functional bioassays involving cell signaling, proliferation, differentiation, migration, and fibrosis studies.

Key validated applications include:

• Cell Proliferation Assays:
  WISP-1 has been used to stimulate proliferation in various cell types, including human and mouse pancreatic beta cells, mesenchymal stem cells, and carcinoma cells. For example, recombinant WISP-1 increased proliferation rates in both mouse and human islets, as measured by Ki67 labeling and insulin co-staining in vitro and in vivo.

• Osteogenic Differentiation Assays:
  Recombinant WISP-1 has been applied to human pluripotent stem cells to promote osteogenic differentiation, as evidenced by increased alkaline phosphatase (ALP) expression and bone marker analysis.

• Angiogenesis and Endothelial Cell Function:
  WISP-1 has been used to treat human coronary artery endothelial cells (HCAECs), where it enhanced proangiogenic and prosurvival genotypes, assessed by Western blot for VEGF-A, Rho-A, RAC, and phospho-BAD/total BAD ratios.

• Fibroblast Activation and Fibrosis Models:
  In studies of cardiac and hepatic fibrosis, recombinant WISP-1 was used to activate human cardiac fibroblasts (HCFs), inducing a myofibroblast phenotype (α-smooth muscle actin expression) and promoting type I collagen synthesis and processing, as measured by Western blot and phenotypic assays. Similarly, it increased migration of hepatic stellate cells in transwell assays.

• Cell Migration Assays:
  Recombinant WISP-1 increased migration of hepatic stellate cells in collagen-coated transwell migration assays, demonstrating its role in cell motility relevant to liver fibrosis.

• Gene Expression and Signaling Pathway Studies:
  WISP-1 has been used to modulate and study gene expression changes in Wnt and cell cycle pathways in mesenchymal stem cells, as well as to investigate its downstream effects in various signaling contexts using qRT-PCR and Western blotting.

Summary Table: Validated Applications of Recombinant Human WISP-1

Additional Notes:

• Most studies use recombinant WISP-1 in bioassays to probe its functional effects on cell behavior.
• Concentrations and protocols vary by application, but typical concentrations range from low ng/mL to several hundred ng/mL depending on the assay.
• WISP-1 is also used as a standard in ELISA for quantification in some studies, though functional bioassays are the primary validated use.

If you need protocols or more specific details for a particular application, please specify the assay or biological context.

To reconstitute and prepare Recombinant Human WISP-1 protein for cell culture experiments, follow these best-practice steps based on current protocols and technical recommendations:

1. Reconstitution

• Buffer selection: Use sterile PBS (phosphate-buffered saline) or sterile distilled water as the reconstitution buffer, depending on the specific formulation and manufacturer’s instructions. PBS is commonly recommended for WISP-1.
• Concentration: Typical reconstitution concentrations range from 0.1–1.0 mg/mL. For example, one protocol specifies reconstituting at 250 μg/mL in sterile PBS, while others suggest 0.1–1.0 mg/mL in water.
• Procedure:
  • Briefly spin down the vial before opening to collect all lyophilized powder at the bottom.
  • Add the appropriate volume of buffer to achieve the desired concentration.
  • Gently mix by inverting or flicking the tube several times. Do not vortex, as vigorous mixing can denature or aggregate the protein.
  • Allow the protein to dissolve slowly at room temperature for 10–30 minutes with gentle agitation.
  • Ensure complete dissolution before use; WISP-1 may dissolve slowly.

2. Handling and Storage

• Aliquoting: After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage: Store aliquots at -20°C to -80°C under sterile conditions. For long-term storage, -80°C is preferred.
• Carrier protein: If stability is a concern, especially at low concentrations, consider adding a carrier protein (e.g., 0.1% BSA) to prevent adsorption to tube walls.

3. Preparation for Cell Culture

• Dilution: Prior to adding to cell culture, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or PBS. Filter sterilize if necessary.
• Avoid foaming: Mix gently to avoid foaming, which can denature the protein.
• Use promptly: Use freshly prepared working solutions for best activity. If not used immediately, keep on ice and avoid prolonged storage at room temperature.

4. Additional Notes

• Do not vortex at any stage, as this can destabilize WISP-1.
• Check solubility: If the protein is slow to dissolve, allow more time at room temperature and gently mix; do not apply excessive mechanical force.
• Sterility: Ensure all buffers and solutions are sterile to prevent contamination in cell culture experiments.

Summary Table: Key Steps for Recombinant Human WISP-1 Preparation

These steps ensure optimal solubility, stability, and biological activity of recombinant WISP-1 for cell culture applications. Always consult the specific product datasheet for any unique requirements.