Betacellulin, also known as BTC, a member of the EGF family of cytokines. BTC is initially synthesized as a glycosylated 32 kDa transmembrane precursor protein, which is processed by proteolytic cleavage to produce the mature sequence (1). Several tissues including kidney, liver, pancreas, uterus, small intestine as well as certain tumor cells express BTC (2). In addition, it can also be found in bodily fluids including serum, milk and colostrum (3). BTC is a potent mitogen for retinal pigment epithelial cells, vascular smooth muscle cells and fibroblasts (4). It also promotes pancreatic beta-cell growth and differentiation (5). The primary receptor for BTC is the receptor for EGF, specifically ErbB-1 and ErbB-4 (6).
Protein Details
Purity
>97% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<0.01EU/µg as determined by the LAL method
Biological Activity
The biological activity of Mouse BTC was determined by its ability to stimulate <sup>3</sup>H-thymidine incorporation by Balb/3T3 mouse fibroblasts. The expected ED<sub>50</sub> is typically 0.1 - 0.4 ng/mL.
The predicted molecular weight of Recombinant Mouse Betacellulin is Mr 9.9 kDa.
Predicted Molecular Mass
9.9
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives.
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
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Recombinant Mouse Betacellulin is used in research applications because it is a potent growth factor that regulates cell proliferation, differentiation, and survival, particularly in pancreatic β-cells, neural stem cells, epithelial cells, and fibroblasts. Its recombinant form ensures high purity, batch-to-batch consistency, and species specificity for mouse models.
Key scientific applications and rationale include:
Pancreatic β-cell research: Betacellulin promotes the growth, differentiation, and neogenesis of pancreatic β-cells, making it valuable for studies on diabetes, islet regeneration, and β-cell development. It has been shown to ameliorate glucose intolerance in diabetic mouse models by inducing β-cell neogenesis.
Cell proliferation and differentiation: Betacellulin is a member of the EGF family and acts as a mitogen for various cell types, including epithelial, neural, and fibroblast cells. It is used to stimulate proliferation in neural stem cell cultures and to prevent spontaneous differentiation.
Tissue regeneration and repair: Betacellulin supports regeneration in tissues such as peripheral nerves by promoting Schwann cell migration and axon elongation, suggesting applications in regenerative medicine and nerve injury models.
Cancer and fibrosis research: Betacellulin is implicated in tumor biology and fibrosis, as it is expressed in several cancer types and can promote proliferation of hepatic stellate cells, contributing to fibrosis and tumor progression.
Bioassays and ELISA standards: Recombinant Betacellulin is validated for use in bioactivity assays and as a standard in ELISA, ensuring reproducibility and reliability in quantitative studies.
Additional considerations:
Species specificity: Using recombinant mouse Betacellulin ensures compatibility with mouse cells and tissues, reducing cross-species variability.
Carrier-free and carrier-added formats: Availability in both formats allows flexibility for different experimental needs, such as avoiding carrier proteins in sensitive assays.
In summary, recombinant mouse Betacellulin is a versatile tool for investigating cell signaling, regeneration, diabetes, cancer, and tissue repair in mouse models, offering high activity and reproducibility for a range of experimental protocols.
Yes, Recombinant Mouse Betacellulin can be used as a standard for quantification or calibration in ELISA assays, provided it is of sufficient purity and properly validated for your assay format.
For ELISA quantification, the standard curve is typically generated using known concentrations of the target protein—either recombinant or purified from natural sources. Recombinant Mouse Betacellulin is commonly supplied for this purpose in commercial ELISA kits and as standalone protein standards. It is critical that the recombinant protein matches the epitope recognized by the antibodies used in your ELISA, and that its concentration is accurately determined.
Key considerations for using recombinant Betacellulin as an ELISA standard:
Purity and Activity: The recombinant protein should be highly pure (typically >80% or higher) and biologically active, as impurities or misfolded protein can affect quantification accuracy.
Validation: Confirm that the recombinant Betacellulin is compatible with your ELISA antibodies (capture and detection), as some antibodies may recognize only specific isoforms or post-translational modifications.
Standard Preparation: Prepare serial dilutions of the recombinant protein in the same buffer as your samples to generate a standard curve. This curve is essential for accurate quantification of unknowns.
Carrier Proteins: Some recombinant standards are supplied with carrier proteins (e.g., BSA) to improve stability and recovery. Carrier-free preparations are preferred if you need to avoid interference in sensitive assays.
Best Practices:
Always verify the concentration of your recombinant standard using an independent method (e.g., absorbance at 280 nm, BCA assay).
Run the standard curve in parallel with your samples on each ELISA plate to account for plate-to-plate variability.
If using a recombinant fragment, ensure it contains the relevant epitopes for antibody recognition.
Summary: Recombinant Mouse Betacellulin is suitable as an ELISA standard if it is pure, correctly quantified, and validated for your assay system. This approach is widely used in both commercial kits and custom ELISA development for accurate quantification of Betacellulin in biological samples.
Recombinant Mouse Betacellulin has been validated for several applications in published research, primarily in bioactivity assays, cell differentiation studies, and functional assays involving cell proliferation, fibrosis, and inflammation.
Key validated applications include:
Bioactivity assays: Recombinant Mouse Betacellulin is routinely validated for bioactivity, such as its ability to stimulate cell proliferation and differentiation in various cell types, including pancreatic beta cells, hepatic stellate cells, and fibroblasts.
Cell differentiation: It has been used to induce differentiation of pancreatic acinar cell lines (e.g., AR42J) into insulin-secreting cells, supporting its role in beta-cell neogenesis and potential diabetes research.
Functional assays: The protein has been validated in functional assays to study its effects on cell signaling pathways, such as EGFR activation, TGF-β2 induction, and integrin upregulation in macrophages, which are relevant to inflammation and fibrosis models.
In vivo studies: Recombinant Mouse Betacellulin has been used in mouse models to promote beta-cell regeneration and ameliorate glucose intolerance, as well as to investigate its role in liver fibrosis and cancer progression.
ELISA and Western blot: Some commercial preparations have been validated for use as standards or controls in ELISA and Western blot applications.
Published research examples:
Liver disease and fibrosis: Betacellulin was shown to promote proliferation of hepatic stellate cells, induce TGF-β2, and increase collagen production, contributing to fibrosis in nonalcoholic steatohepatitis (NASH) models.
Beta-cell neogenesis: Recombinant Betacellulin has been used to promote the formation of new pancreatic beta cells and improve glucose tolerance in diabetic mouse models.
Inflammation and immune modulation: In combination with TLR2/4 agonists, Betacellulin upregulated integrins in macrophages, potentiating inflammatory responses.
In summary: Recombinant Mouse Betacellulin is validated for bioactivity and functional assays in cell culture and animal models, particularly for studies of cell proliferation, differentiation, fibrosis, inflammation, and as a standard in immunoassays.
To reconstitute and prepare Recombinant Mouse Betacellulin protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS. If your preparation is carrier-free, use only sterile PBS; if the formulation contains BSA or you wish to enhance stability, use sterile PBS containing at least 0.1% human or bovine serum albumin.
Step-by-step protocol:
Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
Add sterile PBS (or PBS + 0.1% BSA for carrier-containing formulations) to achieve a final concentration of 100 μg/mL.
Gently mix by swirling or inverting; avoid vigorous shaking to prevent foaming and protein denaturation.
Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
Aliquot the solution to minimize freeze-thaw cycles.
Storage: After reconstitution, store aliquots at –20 °C to –70 °C for long-term use, or at 2–8 °C for up to 1 month under sterile conditions. Avoid repeated freeze-thaw cycles.
Additional notes:
For cell culture, using a carrier protein (BSA or HSA) is recommended to improve stability and prevent adsorption to plasticware.
If your protocol requires a different working concentration, dilute the stock solution further in cell culture medium immediately before use.
Always check the Certificate of Analysis or product datasheet for batch-specific instructions, as formulation details may vary.
Summary Table:
Formulation Type
Reconstitution Buffer
Final Concentration
Carrier Protein Recommended?
Carrier-free
Sterile PBS
100 μg/mL
Optional
With carrier (BSA/HSA)
Sterile PBS + ≥0.1% BSA/HSA
100 μg/mL
Yes
This protocol ensures optimal solubility and biological activity of Betacellulin for cell culture applications.
References & Citations
1. Tada, H. et al. (1999) J. Cell Biochem. 72:423
2. Shing, Y. et al. (1993) Science 259:1604
3. Bastian, SE. et al. (2001) J. Endocrnol. 168:203
4. Dunbar, AJ. et al. (2000) J. Biochem. Cell Bio. 32:805
5. Li, L. et al. (2001) Endocrinology 142:5379
6. Watanabe, T. et al. (1994) J. Bio. Chem. 269:9966