Recombinant Human Betacellulin

Recombinant Human Betacellulin is widely used in research due to its potent biological activities, particularly its role as a growth factor in the EGF (epidermal growth factor) family, making it valuable for studies in cell proliferation, regenerative medicine, diabetes, cancer biology, and stem cell research.

Key reasons to use recombinant human Betacellulin in research applications:

• Potent Mitogenic Activity: Betacellulin is a strong mitogen (stimulator of cell division) for various cell types, including fibroblasts, pancreatic β-cells, retinal pigment epithelial cells, vascular smooth muscle cells, and neural stem cells. This makes it useful for studying cell proliferation, tissue regeneration, and wound healing.

• Beta-Cell Neogenesis and Diabetes Research: Recombinant human Betacellulin has been shown to promote the formation of new pancreatic β-cells and improve glucose tolerance in diabetic animal models, supporting its use in diabetes and pancreatic regeneration studies.

• Cancer and Cell Signaling Studies: As a ligand for the EGF receptor (EGFR), Betacellulin is involved in signaling pathways relevant to cancer cell biology, making it a tool for investigating tumor growth, metastasis, and EGFR-related signaling mechanisms.

• Stem Cell and Neural Research: Betacellulin induces neural stem cell proliferation and helps maintain their undifferentiated state in culture, which is important for neural development and regenerative medicine research.

• Biochemical and Biophysical Studies: Recombinant forms allow for controlled, reproducible experiments, including binding studies (e.g., to heparin or heparan sulfate), receptor activation assays, and structural analyses.

• Assay Development and Standardization: Recombinant Betacellulin is used as a standard in ELISA, Western blotting, and cell-based bioassays due to its defined activity and purity.

• Carrier-Free and Animal-Free Options: Recombinant preparations are available in carrier-free and animal-free formats, reducing the risk of interference or contamination in sensitive applications such as mass cytometry, spatial biology, and cell therapy development.

In summary, recombinant human Betacellulin is a versatile and well-characterized growth factor that enables precise, reproducible studies in cell biology, regenerative medicine, diabetes, cancer, and stem cell research, thanks to its defined activity, purity, and compatibility with a wide range of experimental systems.

Yes, recombinant human Betacellulin can generally be used as a standard for quantification or calibration in ELISA assays, provided it matches the form of Betacellulin detected by your assay. Many commercial ELISA kits for Betacellulin use recombinant human Betacellulin as their calibration standard.

Key considerations and supporting details:

• Recombinant Standard Use: Recombinant human Betacellulin is commonly supplied as the standard in ELISA kits, and protocols often instruct users to prepare a dilution series from this standard for the calibration curve. This practice is standard in quantitative ELISA workflows.

• Formulation: It is recommended to use the recombinant protein with a carrier protein (such as BSA) for ELISA standards, as this enhances stability and reproducibility. Carrier-free formulations are available if BSA or other carriers would interfere with your assay.

• Assay Compatibility: Ensure that the recombinant Betacellulin you use as a standard is structurally and immunologically equivalent to the endogenous Betacellulin detected by your assay. Most ELISA kits are validated for both natural and recombinant forms. However, some kits specifically caution against using recombinant proteins as standards if their epitopes or modifications differ from the native protein in your samples. Always check your kit’s documentation for such restrictions.

• Calibration Curve Preparation: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant standard to generate a reliable calibration curve. Typical standard curves range from low pg/mL to ng/mL concentrations, depending on assay sensitivity.

• Bioactivity and Validation: Recombinant Betacellulin used as a standard is typically validated for bioactivity and purity, ensuring consistent quantification.

Summary Table: Use of Recombinant Human Betacellulin as ELISA Standard

In summary: If your ELISA kit or protocol is validated for recombinant Betacellulin, and the recombinant protein matches the form detected in your samples, you can use it as a standard for quantification or calibration in your ELISA assays. Always verify compatibility with your specific assay system.

Recombinant Human Betacellulin has been validated in published research for several key applications, primarily in studies of cell proliferation, regenerative medicine, diabetes, skin barrier function, fibrosis, and cancer.

Validated Applications:

• Cell Proliferation Assays: Betacellulin is a potent mitogen validated for stimulating proliferation in various cell types, including fibroblasts, retinal pigment epithelial cells, vascular smooth muscle cells, pancreatic α- and β-cells, and neural stem cells.

• Regenerative Medicine and Diabetes Research:

  • Promotes neogenesis and regeneration of pancreatic β-cells, improving glucose tolerance in diabetic animal models.
  • Induces α-cell proliferation and contributes to β-cell regeneration via ErbB3 and ErbB4 signaling pathways.
  • Used in studies investigating therapeutic strategies for insulin-dependent diabetes.

• Skin Barrier Function and Dermatology:

  • Validated for improving skin barrier integrity in models of atopic dermatitis (AD) and Th2 cytokine-mediated skin dysfunction.
  • Enhances expression of tight junction proteins and antimicrobial peptides in human keratinocytes, improving transepithelial electrical resistance (TER) and barrier function.

• Fibrosis and Inflammation:

  • Used to study macrophage activation and integrin upregulation, potentiating inflammation and fibrosis in liver disease models.
  • Investigated as a target for nonalcoholic steatohepatitis (NASH) and liver cancer prevention.

• Cancer Research:

  • Studied for its role in tumorigenesis, particularly in pancreatic β-cell tumors and as a potential therapeutic target in liver cancer.

• Retinal Cell Biology:

  • Promotes proliferation and differentiation of retinal progenitor cells into retinal neurons, relevant for retinal regeneration studies.

• Vascular Biology:

  • Induces DNA synthesis and upregulation of cyclin D1 in human aortic smooth muscle cells, supporting vascular cell proliferation research.

Experimental Protocols and Models:

• In vitro: Used in cell culture systems to stimulate proliferation, differentiation, and barrier function in keratinocytes, fibroblasts, retinal progenitor cells, and pancreatic islet cells.
• In vivo: Applied in animal models of diabetes (alloxan- or streptozotocin-induced), liver fibrosis, and skin disease to assess therapeutic effects and tissue regeneration.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed studies and experimental protocols, confirming the utility of recombinant human betacellulin in diverse fields of biomedical research.

To reconstitute and prepare Recombinant Human Betacellulin protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) or sterile distilled water to a final concentration of 100–500 μg/mL for carrier-free formulations, or 0.1–1.0 mg/mL if using a carrier protein such as 0.1% BSA.

Step-by-step protocol:

• Before opening: Briefly centrifuge the vial to ensure all lyophilized powder is at the bottom.
• Reconstitution:
  • For carrier-free protein, add sterile PBS to achieve 100–500 μg/mL.
  • For formulations requiring carrier protein, use sterile distilled water or PBS containing 0.1% BSA, aiming for 0.1–1.0 mg/mL.
• Mixing: Gently agitate or swirl the vial at room temperature for 15–30 minutes until fully dissolved. Avoid vigorous shaking or vortexing to prevent foaming and protein denaturation.
• Aliquoting: Prepare small aliquots to avoid repeated freeze-thaw cycles, which can degrade protein activity.
• Storage:
  • Store lyophilized protein at –20 °C or lower until use.
  • After reconstitution, store aliquots at –20 °C to –80 °C for long-term storage, or at 2–8 °C for up to 1 month for short-term use.
• Working concentration: For cell culture assays, Betacellulin is typically active at 0.1–1.5 ng/mL in proliferation assays, so dilute the stock solution accordingly in your culture medium.

Additional notes:

• If the protein concentration after reconstitution is below 0.5 mg/mL, adding a carrier protein (e.g., BSA at 0.1%) is recommended to stabilize the protein and prevent adsorption to tube walls.
• Always use sterile technique to prevent contamination.
• Avoid repeated freeze-thaw cycles, as these can reduce protein bioactivity.

Summary Table:

This protocol ensures optimal solubility, stability, and bioactivity of recombinant Betacellulin for cell culture applications.