Recombinant Human FGFR4

Using Recombinant Human FGFR4 in research applications enables precise investigation of the biological functions, signaling mechanisms, and therapeutic targeting of the FGFR4 receptor, which is implicated in diverse physiological and pathological processes including cancer, metabolism, tissue repair, and development.

Key scientific reasons to use recombinant FGFR4 include:

• Cancer Research and Therapeutic Targeting: FGFR4 is a critical mediator in the FGF19-FGFR4 signaling axis, which is frequently dysregulated in cancers such as hepatocellular carcinoma, colon cancer, and ovarian cancer. Recombinant FGFR4 allows for:

  • Screening and characterization of small molecule inhibitors or antibodies targeting FGFR4.
  • Elucidation of downstream signaling pathways (e.g., Ras-Raf-MAPK, GSK3β-Nrf2) upon ligand binding or inhibition.
  • Functional studies to assess the impact of FGFR4 activation or inhibition on cell proliferation, migration, and epithelial-mesenchymal transition (EMT).

• Metabolic and Physiological Studies: FGFR4 regulates bile acid homeostasis, cholesterol metabolism, and muscle differentiation. Recombinant FGFR4 can be used to:

  • Model receptor-ligand interactions (e.g., with FGF19) in vitro.
  • Study receptor-mediated signaling in metabolic tissues.
  • Investigate the effects of genetic variants (such as the Arg388 SNP) on receptor function and stability.

• Tissue Repair and Regenerative Medicine: FGFR4 agonists and recombinant proteins are valuable for exploring roles in tissue repair, regeneration, and protection against tissue damage (e.g., in liver injury models).

• Biochemical and Structural Analysis: Recombinant FGFR4 provides a defined, controllable system for:

  • Structural studies (e.g., crystallography, binding assays).
  • Biochemical assays to dissect receptor activation, phosphorylation, and adaptor protein recruitment.

• Developmental Biology: FGFR4 is essential in embryonic development, angiogenesis, and tissue differentiation. Recombinant protein enables mechanistic studies in developmental models.

• Assay Development and High-Throughput Screening: Recombinant FGFR4 is used to develop and validate assays for drug discovery, including high-throughput screening of FGFR4 modulators.

In summary, recombinant human FGFR4 is a versatile tool for dissecting receptor biology, modeling disease mechanisms, and advancing therapeutic discovery in areas where FGFR4 plays a pivotal role.

Yes, you can use Recombinant Human FGFR4 as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is suitable for your specific ELISA kit and detection system.

Key Points:

• Recombinant FGFR4 as Standard: Recombinant Human FGFR4 proteins (such as those with His-tag or Fc-chimera, e.g., from Abcam, R&D Systems, or other suppliers) are commonly used as standards in ELISA assays to generate a standard curve for quantifying native FGFR4 in samples. These proteins are well-characterized and can be serially diluted to create a reference curve.

• Compatibility: Ensure that the recombinant FGFR4 standard is recognized by the antibodies used in your ELISA kit. Most commercial ELISA kits are validated to detect both native and recombinant FGFR4, but always check the kit’s datasheet or manual for confirmation.

• Formulation: For ELISA standards, it is generally recommended to use recombinant protein without carrier proteins (e.g., BSA) to avoid interference. Carrier-free formulations are preferred for accurate quantification.

• Guidelines: When preparing the standard curve, use serial dilutions of the recombinant FGFR4 in the same buffer as your samples to minimize matrix effects. Follow the manufacturer’s instructions for dilution and assay protocol.

References from Search Results:

• Abcam’s Recombinant Human FGFR-4 Protein Standard (His tag) is suitable for sELISA and SDS-PAGE.
• R&D Systems recommends using recombinant FGFR4 (with or without BSA) as an ELISA standard, with carrier-free versions preferred for quantification.

In summary, yes, Recombinant Human FGFR4 can be used as a standard for ELISA quantification, but ensure compatibility with your kit and use appropriate formulation and dilution protocols.

Recombinant Human FGFR4 has been validated for several key applications in published research, primarily in the context of cancer biology, cell signaling, and therapeutic development.

Validated Applications in Published Research:

• Kinase Assays: Recombinant FGFR4 is frequently used as an active kinase in in vitro kinase assays to study its enzymatic activity, substrate specificity, and inhibition by small molecules or antibodies.
• ELISA (Enzyme-Linked Immunosorbent Assay): It serves as a standard or capture antigen in ELISA to quantify FGFR4 levels or to screen for FGFR4-binding antibodies.
• Cellular Signaling Studies: Recombinant FGFR4 is used to dissect FGF19-FGFR4 signaling pathways, including downstream effects on cell proliferation, migration, metabolism, and differentiation, particularly in hepatocellular carcinoma and other cancers.
• Drug Discovery and Screening: It is employed in high-throughput screening for small-molecule inhibitors or therapeutic antibodies targeting FGFR4, including the development of antibody-drug conjugates (ADCs) and CAR T cells.
• Protein-Protein Interaction Studies: Recombinant FGFR4 is used to characterize binding interactions with ligands (e.g., FGF19), co-receptors, or therapeutic candidates.
• Structural and Biochemical Characterization: It is utilized for crystallography, biophysical assays, and other structural studies to elucidate the receptor’s conformation and ligand-binding properties.
• Cell-Based Functional Assays: Recombinant FGFR4 can be used to stimulate cells or as a target in cell-based assays to assess receptor-mediated signaling, endocytosis, or cytotoxicity (e.g., in CAR T cell or ADC validation).

Supporting Details and Context:

• In cancer research, recombinant FGFR4 has been pivotal for validating FGFR4 as a therapeutic target, especially in hepatocellular carcinoma, rhabdomyosarcoma, and other FGFR4-expressing tumors.
• It is commonly used in in vitro and in vivo models to assess the efficacy of FGFR4-targeted therapies, including kinase inhibitors, monoclonal antibodies, and engineered cell therapies.
• Recombinant FGFR4 is also used as a control or standard in Western blotting and other immunoassays, although direct validation for Western blot is more commonly reported for FGFR4 antibodies rather than the recombinant protein itself.

Summary Table: Applications of Recombinant Human FGFR4 in Research

These applications are supported by a broad range of published studies, particularly in the context of cancer biology and therapeutic development.

To reconstitute and prepare Recombinant Human FGFR4 protein for cell culture experiments, follow these general best practices based on protocols for similar recombinant FGFR4 preparations:

• Centrifuge the vial briefly (e.g., 1–2 minutes at 3,000 × g) before opening to ensure all lyophilized protein is at the bottom.
• Add sterile distilled water or PBS (pH 7.4) to the vial to achieve your desired stock concentration. A typical reconstitution concentration is 0.1–1.0 mg/mL. For most applications, 100 μg/mL in sterile PBS is recommended.
• Gently mix by pipetting up and down or by gentle swirling. Do not vortex, as this may denature the protein.
• Allow the protein to fully dissolve at room temperature for 10–15 minutes. If necessary, gently invert or tilt the vial to aid dissolution.
• If the protein is difficult to dissolve, you may incubate at 4°C for up to 30 minutes, but avoid prolonged exposure to higher temperatures.

Aliquot and Storage:

• Once reconstituted, aliquot the protein to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –80°C for long-term storage (up to 12 months), or at 4°C for short-term use (up to 1 week).
• Avoid repeated freeze-thaw cycles.

Preparation for Cell Culture:

• Before adding to cell culture, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or buffer containing a carrier protein (e.g., 0.1% BSA) to minimize adsorption to plasticware.
• Filter-sterilize the final working solution if sterility is required and the protein solution is not already sterile-filtered.

Summary Table:

Note: Always consult the specific product datasheet for your recombinant FGFR4, as tag, buffer, and formulation may vary between preparations. If your protein is supplied in a buffer containing stabilizers (e.g., trehalose), reconstitution in sterile water is generally sufficient. For cell-based assays, ensure the final buffer is compatible with your cells.

If you need to prepare the protein for functional assays (e.g., ligand stimulation), confirm that the reconstitution buffer does not contain components toxic to your cells or that may interfere with downstream applications.