Recombinant Mouse FGF R3 (IIIc)

Recombinant Mouse FGF R3 (IIIc) is a valuable tool for research applications focused on growth factor signaling, cellular differentiation, and disease modeling, particularly in contexts where the specific IIIc splice variant of FGFR3 is biologically relevant.

FGFR3 (Fibroblast Growth Factor Receptor 3) is a receptor tyrosine kinase that regulates key processes such as cell growth, division, differentiation, angiogenesis, wound healing, and embryonic development. The IIIc isoform arises from alternative splicing and exhibits distinct ligand-binding specificity compared to other FGFR3 variants, making it crucial for studies where isoform-specific signaling is implicated.

Key scientific applications for Recombinant Mouse FGF R3 (IIIc):

• Growth Factor Signaling Studies: FGFR3-IIIc mediates unique cellular responses to fibroblast growth factors, enabling precise dissection of FGF signaling pathways in mouse models.
• Cancer Research: FGFR3-IIIc has been shown to exert oncogenic effects in certain cancers, such as colorectal cancer, by promoting tumor cell growth and survival. Using the recombinant protein allows for controlled investigation of these mechanisms in vitro and in vivo.
• Developmental Biology: FGFR3 is essential for bone and brain development. The IIIc isoform is particularly relevant for skeletal and neural studies, as its expression and function differ from other splice variants.
• Disease Modeling: Mutations and dysregulation of FGFR3, including the IIIc variant, are linked to skeletal dysplasias and benign tumors. Recombinant FGFR3-IIIc can be used to model these conditions and test therapeutic interventions.
• Cellular Differentiation and Stem Cell Research: FGFR3-IIIc is expressed in neural stem cells and astrocytes, making it useful for studies on CNS development, injury response, and stem cell biology.

Technical advantages:

• Isoform specificity: Recombinant FGFR3-IIIc enables experiments that distinguish between the biological effects of IIIc versus IIIb or other isoforms, which is critical for understanding tissue-specific and disease-specific signaling.
• High purity and activity: Recombinant preparations are typically >90% pure and suitable for ELISA, cell-based assays, and biochemical studies.

In summary, using Recombinant Mouse FGF R3 (IIIc) allows for precise, isoform-specific investigation of FGFR3-mediated signaling in mouse systems, supporting research in cancer biology, developmental processes, and disease modeling where the IIIc variant plays a pivotal role.

You can use recombinant Mouse FGF R3 (IIIc) protein as a standard for quantification or calibration in ELISA assays, provided that the ELISA kit is designed to detect FGFR3 and the recombinant protein matches the epitope recognized by the assay antibodies.

For accurate quantification:

• The standard curve in ELISA should be generated using a purified protein, ideally the same recombinant form as your target analyte.
• Recombinant proteins are commonly used as standards in quantitative ELISA protocols, as long as their purity and concentration are well-characterized.
• The recombinant Mouse FGFR3 (IIIc) protein should be reconstituted and diluted according to the ELISA kit instructions to create a standard curve covering the expected concentration range in your samples.
• Ensure the recombinant protein’s sequence and post-translational modifications (if any) are compatible with the antibodies used in your ELISA, as some kits are validated only for specific isoforms or domains.

Best practices:

• Confirm that your ELISA kit documentation supports the use of recombinant FGFR3 (IIIc) as a standard, as some kits may provide their own validated standard or require a specific isoform.
• Validate the standard curve by running serial dilutions of the recombinant protein and checking for parallelism with endogenous samples, ensuring accurate quantification.
• Use carrier proteins (e.g., BSA) if recommended for stability, but avoid them if the ELISA is sensitive to carrier interference.

Limitations:

• If your recombinant FGFR3 (IIIc) differs in glycosylation or other modifications from the native protein, quantification may be relative rather than absolute.
• Always follow the ELISA kit’s instructions for standard preparation and curve fitting (e.g., 4PL analysis).

In summary, recombinant Mouse FGFR3 (IIIc) is suitable as an ELISA standard if it matches the assay’s specificity and is properly validated for your application.

Recombinant Mouse FGF R3 (IIIc) has been validated for several key applications in published research, primarily related to its role in signaling, cellular assays, and antibody development. Based on the available literature and product information, the main applications include:

• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant Mouse FGF R3 (IIIc) is commonly used as a target antigen in ELISA assays to study antibody binding specificity, particularly for anti-FGFR3 antibodies. Studies have used it to validate the binding of monoclonal and polyclonal antibodies, including those with specificity for the IIIc isoform.

• Cell Panning and Antibody Selection: The protein has been immobilized for use in cell panning and phage display techniques to select and amplify scFv (single-chain variable fragment) binders and antibodies specific for FGFR3-IIIc. This approach is used for generating functional antibodies with biological activity.

• Functional Assays (Cell Viability and Proliferation): Recombinant Mouse FGF R3 (IIIc) is used in assays to assess the biological effects of anti-FGFR3 antibodies, such as inhibition of cell viability and tumor growth in FGFR3-overexpressing cell lines.

• Flow Cytometry and Cell Surface Labeling: The protein has been used to confirm the expression and labeling efficiency of FGFR3 on cell surfaces, often in conjunction with biotinylated probes or fluorescent tags.

• Western Blotting: The protein is suitable for use in Western blot applications to detect FGFR3 expression and to validate antibody specificity.

• Bioactivity Assays: Recombinant Mouse FGF R3 (IIIc) is used to study the biological activity of FGF ligands and their interactions with FGFR3, including downstream signaling and cellular responses.

These applications highlight the utility of Recombinant Mouse FGF R3 (IIIc) in both basic research and antibody development, particularly in studies focused on growth factor signaling, cancer biology, and regenerative medicine.

To reconstitute and prepare Recombinant Mouse FGF R3 (IIIc) protein for cell culture experiments, add sterile water or sterile PBS to the lyophilized protein to achieve a recommended stock concentration, typically between 0.1–1.0 mg/mL. Use aseptic technique throughout, and aliquot the solution to avoid repeated freeze-thaw cycles.

Step-by-step protocol:

• Before opening the vial: Briefly centrifuge to collect all lyophilized powder at the bottom.
• Reconstitution buffer: Most preparations are lyophilized from PBS (pH 7.2–7.3) without calcium or magnesium. If not specified, use sterile, endotoxin-free water or sterile PBS. Check the product datasheet for specific buffer requirements.
• Concentration: Commonly, reconstitute to 0.1–1.0 mg/mL. For example, add 100 µL to 100 µg of protein for 1 mg/mL, or 1 mL for 0.1 mg/mL.
• Mixing: Gently pipette up and down or swirl to dissolve. Avoid vigorous vortexing to prevent protein denaturation.
• Carrier protein: If the protein is to be stored long-term or used at low concentrations, consider adding a carrier protein (e.g., 0.1% BSA) to stabilize the solution.
• Aliquoting: Divide into single-use aliquots to minimize freeze-thaw cycles.
• Storage: Store reconstituted protein at 2–8°C for short-term (up to 1 month), or at –20°C to –80°C for long-term. Avoid repeated freeze-thaw cycles.
• Sterility: Use only sterile reagents and equipment. Filter-sterilize the final solution if necessary.

Preparation for cell culture:

• Dilution: Immediately before use, dilute the stock solution into cell culture medium to the desired working concentration. Ensure the final buffer composition is compatible with your cells (e.g., avoid high salt or additives that may affect cell viability).
• Bioactivity: If required, confirm protein activity by SDS-PAGE or a functional assay, as some preparations may not be validated for bioactivity.

Additional notes:

• If the formulation contains stabilizers (e.g., trehalose, mannitol), these are generally compatible with cell culture but should be considered when preparing working solutions.
• Always consult the specific product datasheet for lot-specific instructions, as formulations and recommended reconstitution buffers may vary.

This protocol ensures optimal solubility, stability, and activity of recombinant mouse FGF R3 (IIIc) for cell culture applications.