Recombinant Mouse TRAIL R2

Using Recombinant Mouse TRAIL R2 in research applications is essential for studying the mechanisms of TRAIL-induced apoptosis, cancer cell signaling, and therapeutic targeting in mouse models. This recombinant protein enables precise investigation of receptor-ligand interactions, apoptotic pathways, and the role of TRAIL R2 in tumor progression and metastasis.

Key reasons to use recombinant mouse TRAIL R2 include:

• Modeling TRAIL-induced apoptosis: TRAIL R2 (also known as TNFRSF10B) is a primary receptor mediating apoptosis upon binding to its ligand, TRAIL. Recombinant forms allow controlled in vitro and in vivo studies of apoptosis in mouse cells, which is critical for understanding cancer cell sensitivity and resistance mechanisms.
• Cancer research and therapy development: TRAIL R2 is implicated in tumor cell proliferation, migration, and metastasis. For example, knockdown of TRAIL R2 in mouse models reduces bone metastasis and tumor proliferation, highlighting its role in cancer progression and as a potential therapeutic target.
• Testing receptor-specific interventions: Recombinant TRAIL R2 can be used to screen and characterize antibodies, small molecules, or other biologics that modulate receptor function, enabling the development of targeted therapies.
• Species-specific studies: Mouse TRAIL R2 is structurally and functionally distinct from its human counterpart, so using the recombinant mouse protein is necessary for accurate preclinical studies in murine systems, including antibody validation, ligand binding assays, and functional analyses.
• Mechanistic studies of metastasis and signaling: TRAIL R2 regulates key signaling pathways (e.g., HMGA2, p-Src, p-Akt, CXCR4) involved in cell proliferation, migration, and epithelial-mesenchymal transition, making it a valuable tool for dissecting these processes in mouse models.

In summary, recombinant mouse TRAIL R2 is a critical reagent for dissecting apoptosis pathways, evaluating anti-cancer strategies, and modeling disease mechanisms in murine systems, thereby advancing both basic and translational research.

You can use recombinant Mouse TRAIL R2 as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately determined, but several important considerations must be addressed for reliable quantification.

Key considerations:

• Protein Purity and Formulation: The recombinant protein should be highly purified and, ideally, formulated in a buffer compatible with your ELISA system. Carrier-free or BSA-containing formulations may be chosen depending on your application, but carrier-free is generally preferred for standard curve preparation to avoid interference.

• Concentration Assignment: The mass concentration stated on the vial may not always reflect the immunologically detectable amount in your ELISA, due to differences in protein folding, post-translational modifications, or lot-to-lot variability. It is best practice to assign the value of your standard by running it in the ELISA and calibrating it against a reference standard, if available.

• Validation: Before using the recombinant TRAIL R2 as a standard, validate its performance in your specific ELISA setup. This includes confirming that the recombinant protein is recognized by the capture and detection antibodies and that it produces a standard curve with appropriate sensitivity and linearity.

• Documentation and Controls: Document the source, lot, and preparation method of your recombinant standard. Include appropriate controls and, if possible, compare your standard curve to one generated with a commercially validated ELISA kit standard to ensure accuracy.

Best practices for using recombinant proteins as ELISA standards:

• Use a purified recombinant protein with a well-characterized concentration.
• Prepare serial dilutions in the same matrix as your samples to minimize matrix effects.
• Validate the standard curve for linearity, sensitivity, and reproducibility within your assay range.
• If using a recombinant protein not specifically validated as an ELISA standard, assign its value based on its performance in your assay, not solely on the label concentration.

Summary:
Recombinant Mouse TRAIL R2 can be used as a standard for ELISA quantification if it is pure, its concentration is accurately determined, and its performance is validated in your assay. For best results, calibrate it within your ELISA system and do not rely solely on the manufacturer’s stated concentration.

Recombinant Mouse TRAIL R2 (TNFRSF10B) has been validated for several key applications in published research, primarily in the context of apoptosis, cancer biology, and cell signaling studies.

Validated Applications:

• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant Mouse TRAIL R2 is used as a standard or capture protein in ELISA to quantify TRAIL R2 levels or to study ligand-receptor interactions.
• Western Blotting: It serves as a positive control or antigen for antibody validation in Western blot assays, enabling detection and quantification of TRAIL R2 protein expression.
• Cell Culture Studies: Recombinant Mouse TRAIL R2 is applied in cell-based assays to investigate its role in apoptosis, particularly in response to TRAIL ligand stimulation. It is used to study the induction of apoptosis in cancer cell lines and primary cells.
• Functional Assays: The protein is utilized to assess its ability to inhibit TRAIL-mediated cytotoxicity, often by acting as a decoy receptor or by modulating TRAIL signaling pathways in vitro.
• Protein-Protein Interaction Studies: Recombinant TRAIL R2 is used to characterize binding interactions with TRAIL or other ligands, including structural and mechanistic studies.
• Fusion Protein Engineering: It is incorporated into recombinant fusion constructs to enhance therapeutic efficacy or alter signaling properties, especially in the development of novel anticancer agents.

Additional Context:

• Cancer Research: Recombinant TRAIL R2 is extensively used to study mechanisms of TRAIL-induced apoptosis in tumor cells, evaluate resistance mechanisms, and develop TRAIL-based therapeutics.
• Signaling Pathway Analysis: It is employed to dissect downstream signaling events such as activation of caspases, mitochondrial pathways, and non-canonical signaling (e.g., NF-κB, MAPK, PI3K/Akt).
• Clinical Translation: Recombinant TRAIL R2 and its fusion proteins have been tested in preclinical and clinical studies for their potential as anticancer therapeutics, either alone or in combination with other agents.

Summary Table of Validated Applications

These applications are supported by published research and technical documentation, confirming the utility of recombinant Mouse TRAIL R2 in both basic and translational studies focused on apoptosis, cancer therapy, and cell signaling.

To reconstitute and prepare Recombinant Mouse TRAIL R2 protein for cell culture experiments, follow these general steps, which are consistent with best practices for recombinant protein handling:

1. Equilibrate Materials
   Bring the lyophilized protein vial and your reconstitution buffer (typically sterile distilled water or PBS, pH 7.0–7.4) to room temperature before opening to minimize condensation.

2. Centrifuge the Vial
   Briefly centrifuge the vial to collect all lyophilized material at the bottom.

3. Reconstitution

   • Add sterile distilled water or PBS to the vial. The recommended concentration for reconstitution is typically 0.1–1.0 mg/mL.
   • For example, to reconstitute 100 µg of protein, add 100 µL (for 1 mg/mL) to 1 mL (for 0.1 mg/mL) of buffer.
   • Gently swirl or invert the vial to dissolve. Avoid vigorous vortexing, which can denature the protein.
   • If the protein appears as a film or does not dissolve immediately, allow it to sit at room temperature for 15–30 minutes with gentle agitation. If undissolved flakes remain, continue gentle mixing for up to 2 hours.

4. Carrier Protein (Optional but Recommended)
   Adding a carrier protein such as 0.1% BSA can enhance stability and prevent adsorption to plastic surfaces, especially at low concentrations.

5. Aliquoting and Storage

   • Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C to –80°C for long-term storage. For short-term use (up to 1 week), store at 4°C.
   • Avoid multiple freeze-thaw cycles, as this can reduce protein activity.

6. Preparation for Cell Culture

   • Before adding to cell culture, dilute the protein to the desired working concentration using sterile cell culture medium or buffer.
   • Filter-sterilize the final solution if sterility is required and the protein is compatible with filtration.

Additional Notes:

• Always consult the specific product datasheet or Certificate of Analysis for any protein-specific instructions, as some preparations may require unique buffers or additives.
• If the protein is fused to an Fc tag, as is common for TRAIL R2, standard reconstitution protocols apply, but confirm compatibility with your downstream assay.

Summary Table: Recombinant Mouse TRAIL R2 Protein Reconstitution

These guidelines will ensure optimal solubility, stability, and activity of recombinant Mouse TRAIL R2 protein for cell culture applications.