Recombinant Mouse TRAIL

Recombinant Mouse TRAIL (TNF-related apoptosis-inducing ligand) is widely used in research to study apoptosis, tumor immunology, and disease mechanisms in murine models because it specifically interacts with mouse TRAIL receptors, enabling accurate modeling of TRAIL-mediated pathways in mice.

Key reasons to use recombinant mouse TRAIL in research applications:

• Species-Specific Activity: Mouse TRAIL binds effectively to murine TRAIL receptors, unlike human TRAIL, which has weak affinity for mouse receptors. This ensures that experimental results in mouse models accurately reflect TRAIL biology relevant to the species.

• Tumor Suppression and Apoptosis: Recombinant mouse TRAIL induces apoptosis in tumor cells and has been shown to inhibit tumor growth and prolong survival in tumor-bearing mice. This makes it a valuable tool for studying cancer therapies and tumor immune microenvironment modulation.

• Immunomodulatory Effects: TRAIL has dose-dependent immunomodulatory roles. At optimal doses, it activates innate immune cells and CD8^+^ T cells, enhancing antitumor immunity. However, higher doses can promote a tumor-supportive immune environment, highlighting the importance of dose selection in experimental design.

• Disease Modeling Beyond Cancer: Recombinant mouse TRAIL is also used to study non-cancer diseases, such as pulmonary arterial hypertension (PAH), where it has been shown to mediate vascular remodeling and disease progression in rodent models.

• Mechanistic Studies: Using recombinant mouse TRAIL allows for mechanistic dissection of apoptosis, immune cell activation, and cytokine signaling pathways in genetically modified or wild-type mice.

• Preclinical Therapeutic Evaluation: It enables preclinical testing of TRAIL-based therapies, combination regimens, and immune interventions in mouse models before translation to human studies.

• Biomarker Discovery: Mouse models treated with recombinant TRAIL can be used for biomarker discovery, helping to identify molecular signatures of response or resistance to TRAIL-based therapies.

In summary, recombinant mouse TRAIL is essential for accurately modeling TRAIL biology in murine systems, enabling robust studies of apoptosis, immunomodulation, and disease mechanisms relevant to both cancer and non-cancer pathologies.

Yes, recombinant Mouse TRAIL can generally be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common practice in quantitative ELISA protocols for cytokines and other proteins.

Supporting details and best practices:

• Recognition by ELISA kits: Most commercial Mouse TRAIL ELISA kits are designed to detect both natural and recombinant Mouse TRAIL as standards and in samples. The assay antibodies are validated to recognize the recombinant form, ensuring accurate quantification when used as a standard.

• Standard curve preparation: For quantitative ELISA, a standard curve is generated using serial dilutions of a known concentration of the target protein—recombinant Mouse TRAIL in this case. The accuracy of your quantification depends on the purity and correct concentration assignment of your recombinant standard.

• Protein purity and carrier considerations: It is recommended to use a highly purified recombinant protein for standard curve preparation. Some suppliers offer recombinant proteins with or without carrier proteins (e.g., BSA). Carrier-free preparations are preferred for precise quantification, but BSA-containing forms may be used if compatible with your assay.

• Matrix effects: Ideally, the standard should be diluted in the same buffer or matrix as your samples to minimize matrix effects and ensure comparable detection. If your samples are in serum, plasma, or culture medium, consider preparing your standard curve in the same matrix or use the kit’s recommended standard diluent.

• Validation: Always verify that your recombinant Mouse TRAIL is recognized by the specific ELISA kit you are using. Most kits specify compatibility with recombinant standards, but it is good practice to check the kit documentation or perform a pilot experiment.

Summary of best practices:

• Use a highly purified, accurately quantified recombinant Mouse TRAIL.
• Prepare the standard curve according to the ELISA kit instructions.
• Match the standard diluent to your sample matrix when possible.
• Confirm that your ELISA kit detects recombinant Mouse TRAIL equivalently to native protein.

In conclusion, recombinant Mouse TRAIL is suitable as a standard for ELISA quantification, provided these guidelines are followed for optimal accuracy and reproducibility.

Recombinant mouse TRAIL (TNF-related apoptosis-inducing ligand, also known as TNFSF10) has been validated for several important research applications in published studies:

Cancer Cell Apoptosis Studies

Recombinant mouse TRAIL has been extensively validated for inducing apoptosis in cancer cell lines. The protein demonstrates dose-dependent apoptotic activity, with studies showing it can induce apoptosis in TRAIL-sensitive cell lines within relatively short timeframes. Research has utilized recombinant mouse TRAIL in various tumor models, including 4T1 (breast cancer), CT26 (colon cancer), and B16 (melanoma) models, to evaluate antitumor efficacy in vivo.

Immunomodulatory Research

A significant application involves studying the immunomodulatory effects of recombinant TRAIL in tumor microenvironments. Soluble mouse TRAIL (smTRAIL) has been validated for examining dose-related bidirectional regulatory roles in immune responses, including activation of innate immune cells and CD8+ T cells at optimal doses. This research has demonstrated that recombinant mouse TRAIL can modulate tumor-associated macrophages and reshape the tumor immune microenvironment.

Immune Cell Function Studies

Recombinant mouse TRAIL has been validated for investigating effects on immune cell populations. Specifically, it has been used to study inhibition of secondary clonal expansion of mouse CD8+ T cells, making it useful for understanding T cell regulation and immune checkpoint mechanisms.

In Vitro and In Vivo Experimental Models

The protein is suitable for both cell culture applications and animal model studies. It has been validated for use in establishing tumor-bearing mouse models to evaluate therapeutic potential and for examining molecular mechanisms of action through techniques such as Western blotting, gene expression analysis, and immunological detection in tumor tissues.

To reconstitute and prepare Recombinant Mouse TRAIL protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the lyophilized protein in sterile, ultrapure water (18 MΩ-cm H₂O) at a concentration of at least 100 µg/mL. For example, add 100 µL water to 100 µg protein for a 1 mg/mL stock.
• Mix gently by swirling or inverting; avoid vortexing, as vigorous agitation can denature the protein.
• Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
• If the protein does not dissolve completely, repeat gentle mixing and brief centrifugation.
• Dilute the reconstituted stock to your desired working concentration using cell culture medium or buffer. For enhanced stability, especially at low concentrations, add a carrier protein such as 0.1% BSA or HSA.
• Aliquot the solution to avoid repeated freeze-thaw cycles.
• Storage after reconstitution:
  • Short-term (2–7 days): Store at 2–8 °C.
  • Long-term: Store aliquots at –20 °C or below, ideally with carrier protein to prevent loss of activity.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

Additional notes:

• For serum-free or in vivo applications, avoid animal-derived carrier proteins; use alternatives like trehalose if needed.
• Always consult the specific Certificate of Analysis (CoA) for your batch for any unique instructions.

Summary Table: Key Steps for Recombinant Mouse TRAIL Reconstitution

These steps will ensure optimal solubility, stability, and biological activity of recombinant Mouse TRAIL for cell culture experiments.