Recombinant Mouse Ret

Recombinant Mouse Ret is used in research applications to study the function, signaling, and disease relevance of the RET receptor tyrosine kinase, particularly in mouse models of development, cancer, and neurobiology. It provides a controlled, reproducible tool for dissecting RET-mediated pathways and for screening potential therapeutics targeting RET-driven diseases.

Key scientific reasons to use recombinant Mouse Ret include:

• Functional Studies: Recombinant Mouse Ret enables precise investigation of RET’s role in cell signaling, neuronal development, and oncogenesis. For example, recombinant RET proteins have been shown to induce T-lymphocyte proliferation and modulate immune responses in mouse models, which is relevant for immunology and cancer research.

• Disease Modeling: RET is implicated in several cancers (e.g., medullary thyroid carcinoma, lung adenocarcinoma) and developmental disorders. Using recombinant Mouse Ret allows researchers to model RET-driven diseases in vitro and in vivo, facilitating the study of disease mechanisms and the evaluation of targeted therapies.

• Drug Screening and Mechanistic Assays: Recombinant Mouse Ret is essential for high-throughput screening of RET inhibitors and for biochemical assays to characterize inhibitor specificity, potency, and mechanism of action. For instance, studies have used recombinant RET to demonstrate the selectivity and efficacy of next-generation RET inhibitors against wild-type and mutant forms of the protein.

• Protein-Protein Interaction Studies: The recombinant protein can be used in binding assays to identify and characterize interactions with ligands, co-receptors, or therapeutic antibodies, supporting both basic research and drug development.

• Reproducibility and Standardization: Recombinant proteins offer batch-to-batch consistency, defined purity, and controlled post-translational modifications, which are critical for generating reproducible and interpretable experimental data.

• Compatibility with Mouse Models: Using the mouse version of RET ensures species compatibility in mouse-based experiments, reducing immunogenicity and improving physiological relevance compared to using human RET in mouse systems.

In summary, recombinant Mouse Ret is a versatile and essential reagent for dissecting RET biology, modeling RET-associated diseases, and advancing therapeutic discovery in mouse-based research systems.

Yes, you can use Recombinant Mouse Ret as a standard for quantification or calibration in your ELISA assays, provided that the ELISA kit or antibody pair you are using is designed to detect mouse Ret. Recombinant proteins, such as Recombinant Mouse Ret, are commonly used as standards in ELISA to generate a standard curve for quantifying the target analyte in your samples.

However, there are several important considerations:

1. Specificity: Ensure that the recombinant protein is recognized by the antibodies in your ELISA system. The recombinant protein should match the epitope(s) targeted by your assay.

2. Purity and Concentration: Use a highly purified recombinant protein, and accurately determine its concentration (e.g., by amino acid analysis, HPLC, or spectrophotometry). The concentration stated on the vial may not always reflect the immunologically active amount, so it is best to value-assign the standard based on its performance in your ELISA.

3. Matrix Effects: For complex sample types (e.g., serum, plasma), it is recommended to include a spike control—dilute the standard in the same matrix as your samples to check for matrix interference and ensure accurate quantification.

4. Standard Curve Range: Prepare a dilution series of the recombinant standard that covers the expected concentration range of your samples. The standard curve should be generated in each assay to ensure accurate quantification.

5. Validation: Validate the use of the recombinant standard in your specific ELISA protocol by checking recovery and linearity. Acceptable recovery is typically 80–120%.

In summary, Recombinant Mouse Ret can be used as a standard for ELISA quantification, but it must be validated for your specific assay conditions and sample type.

Recombinant Mouse Ret has been validated for several key applications in published research, primarily in studies of cell signaling, cancer biology, and drug screening.

Validated Applications:

• Bioassays: Recombinant Mouse Ret is frequently used in bioactivity assays to study RET-mediated signaling pathways, receptor activation, and downstream effects in various cell types.
• Cell Culture: It is applied in cell culture systems, especially for generating and characterizing murine RET fusion oncogenes and evaluating RET-dependent cellular responses.
• In Vivo Models: Recombinant Mouse Ret has been utilized in mouse models to assess the efficacy of RET inhibitors, investigate RET-driven tumor growth, and study brain metastasis in RET-driven cancers.
• Drug Screening: It is used to evaluate the potency and selectivity of RET inhibitors against wild-type and mutant RET kinases, including resistance-associated mutations (e.g., V804, G810).
• Functional Studies: Research includes rational design of heterodimeric receptors and analysis of RET’s role in cell survival, differentiation, and disease models.

Supporting Details:

• Cancer Research: RET is a critical target in studies of lung, thyroid, colon, and pancreatic cancers, especially those driven by RET fusions or mutations. Recombinant Mouse Ret enables preclinical testing of selective inhibitors and assessment of tumor control in xenograft and orthotopic models.
• Signaling Pathways: RET’s role as a receptor tyrosine kinase makes recombinant protein essential for dissecting signaling mechanisms and validating therapeutic strategies.
• Resistance Mechanisms: Recombinant Mouse Ret is used to model and overcome resistance mutations in RET, facilitating the development of next-generation inhibitors.

Summary Table of Applications

These applications are supported by multiple peer-reviewed studies, demonstrating the versatility and importance of recombinant Mouse Ret in both basic and translational research.

To reconstitute and prepare Recombinant Mouse Ret protein for cell culture experiments, dissolve the lyophilized protein in sterile water or sterile PBS to achieve a stock concentration typically between 0.1–1.0 mg/mL. The exact diluent and concentration should be confirmed with the product’s Certificate of Analysis or datasheet, but these ranges are standard for recombinant proteins used in cell culture.

Step-by-step protocol:

• Reconstitution:

  • Add sterile water or sterile PBS directly to the vial containing the lyophilized protein.
  • For most applications, a concentration of 100 μg/mL is recommended (e.g., add 1 mL to 100 μg protein).
  • Gently mix by pipetting or slow inversion; avoid vigorous shaking or vortexing to prevent protein denaturation and foaming.
  • Allow the protein to fully dissolve at room temperature for 15–30 minutes.

• Aliquoting and Storage:

  • Once reconstituted, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
  • Store aliquots at –20°C to –80°C for long-term preservation.
  • Use freshly thawed aliquots for experiments and avoid storing working solutions for more than one week at 4°C.

• Preparation for Cell Culture:

  • Dilute the stock solution into your cell culture medium to the desired final concentration, typically in the range of ng/mL to μg/mL, depending on experimental requirements.
  • Ensure the final buffer composition is compatible with your cells (e.g., avoid high salt or additives that may affect cell viability).
  • Filter the working solution through a 0.22 μm filter if sterility is required.

Additional notes:

• If the protein is sensitive or prone to aggregation, consider adding carrier proteins (e.g., 0.1% BSA) or glycerol (5–50%) to stabilize the solution.
• Always consult the specific product datasheet for recommended reconstitution conditions, as some proteins may require special buffers or additives.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant Mouse Ret protein for cell culture experiments.