Recombinant Mouse IL-16

Recombinant Mouse IL-16 is used in research applications to study and manipulate immune responses, particularly those involving T cell activation, macrophage polarization, and inflammation in murine models. Its recombinant form allows for controlled, reproducible experiments in immunology, oncology, and inflammation research.

Key scientific applications and rationale include:

• Enhancing Anti-Tumor Immunity: Recombinant mouse IL-16 administration in cancer models promotes a Th1-dominant tumor microenvironment by reprogramming glutamine metabolism in CD4⁺ T cells, increasing IFN-γ production, and facilitating Th1 cell-macrophage crosstalk. This leads to improved anti-tumor immune responses and greater efficacy of immunotherapies such as immune checkpoint blockade.

• Modulating Immune Cell Function: IL-16 acts as a chemoattractant for CD4⁺ T cells and modulates T cell activation, making it valuable for studying T cell recruitment, differentiation, and cytokine production in vitro and in vivo. It also influences macrophage polarization toward an M1-like phenotype, which is associated with pro-inflammatory and anti-tumor activity.

• Investigating Inflammatory Pathways: Active IL-16 drives the production of proinflammatory cytokines (IL-6, TNF, IL-1β) by peripheral blood mononuclear cells and is implicated in various inflammatory diseases, including autoimmune disorders and tissue injury models. Recombinant IL-16 enables mechanistic studies of these pathways.

• Cell Proliferation and Growth Assays: Recombinant mouse IL-16 can stimulate cell growth in certain cell lines, such as Jurkat T lymphoblastoid cells, making it useful for proliferation assays and functional studies of lymphocyte biology.

• Standardization in Detection and Quantification: Recombinant IL-16 serves as a standard for ELISA and other immunoassays, facilitating the quantification of endogenous IL-16 in biological samples and the screening of antibodies or inhibitors targeting IL-16 signaling.

• Modeling Disease Mechanisms: IL-16 has been shown to impair muscle function and contribute to muscle atrophy in aging models, and its recombinant form is used to dissect these effects and test therapeutic interventions.

Best practices:

• Use recombinant mouse IL-16 for species-specific studies to ensure biological relevance and avoid cross-reactivity issues seen with human IL-16.
• Employ recombinant proteins for reproducibility, defined activity, and purity in mechanistic and therapeutic research.

In summary, recombinant mouse IL-16 is a versatile tool for dissecting immune mechanisms, modeling disease, and developing immunotherapeutic strategies in murine systems.

Yes, you can use recombinant Mouse IL-16 as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is highly purified and its concentration is accurately known.

Key Points to Consider:

• Purity and Quality: The recombinant Mouse IL-16 should be >98% pure (as confirmed by SDS-PAGE or other methods) and free of contaminants that could interfere with the assay. For example, yeast-derived recombinant Mouse IL-16 is described as naturally endotoxin-free and suitable for use as an ELISA standard.

• Quantitative Use: To generate a reliable standard curve, the recombinant protein must be accurately quantified (e.g., by amino acid analysis, HPLC, or spectrophotometry). This allows you to prepare a series of dilutions with known concentrations for your ELISA.

• Compatibility: The recombinant IL-16 should be compatible with the detection antibodies and assay conditions used in your ELISA. Most commercial ELISA kits for Mouse IL-16 include a recombinant standard, confirming that recombinant IL-16 is appropriate for this purpose.

• Guidelines: Best practice is to use a purified or recombinant protein for the standard curve to ensure accurate quantification. If you are using a kit, follow the manufacturer’s instructions for preparing the standard curve.

References from Search Results:

• Result states: "Mouse IL-16 applications are for cell culture, ELISA standard, and Western Blot Control."
• Result recommends: "A purified protein should be used to prepare the standard curve. Otherwise, use a recombinant protein which can be semi-purified in the lab and measure the concentration with HPLC."
• Result and mention that ELISA kits for Mouse IL-16 include a recombinant standard for generating the standard curve.

In summary, recombinant Mouse IL-16 is suitable for use as a standard in ELISA assays, as long as it is highly purified and accurately quantified.

Validated Applications in Published Research

Recombinant mouse IL-16 has been validated for several significant applications in cancer immunotherapy and immune research:

Anti-tumor Immune Response Enhancement

The primary validated application involves enhancing anti-tumor immunity through multiple mechanisms. Recombinant IL-16 administration has been demonstrated to establish a Th1-dominant tumor microenvironment (TME) by inhibiting glutamine catabolism through downregulation of glutaminase in CD4+ T cells, thereby increasing production of the Th1 effector cytokine interferon-gamma (IFN-γ). In murine breast cancer (E0771) and lung cancer (LLC) models, mice treated with recombinant IL-16 developed significantly smaller tumors compared to control mice. Additionally, recombinant IL-16 significantly reduced the growth of spontaneously developed mammary tumors in transgenic mouse models.

Immune Checkpoint Blockade Potentiation

Recombinant IL-16 has been validated as an adjuvant to enhance the effectiveness of immune checkpoint blockade (ICB) therapy. The cytokine augments expression of CXCR3 ligands (CXCL9 and CXCL10) in tumor-associated macrophages, which are required for optimal anti-tumor immune responses following ICB. Mice treated with anti-PD-L1 antibody combined with recombinant IL-16 showed significantly higher levels of IFN-γ, perforin, and granzyme B in the TME compared to ICB monotherapy.

Tumor-Associated Macrophage Reprogramming

Recombinant IL-16 has been validated for its ability to reprogram tumor-associated macrophages toward an M1-like pro-inflammatory phenotype through Th1 cell-macrophage crosstalk, contributing to the overall anti-tumor immune response.

Assay Development and Detection

Recombinant mouse IL-16 serves as a standard for IL-16 detection and quantification assays, as well as for screening and release assays for antibodies that block IL-16 signaling.

To reconstitute and prepare Recombinant Mouse IL-16 protein for cell culture experiments, follow these best-practice steps:

• Before opening the vial:

  • Briefly centrifuge the vial (e.g., 3000 rpm for 5 minutes or 20–30 seconds in a microcentrifuge) to ensure all lyophilized protein is at the bottom and not on the cap or sides.

• Reconstitution:

  • Use sterile deionized or distilled water as the solvent unless your protocol specifies a different buffer.
  • Recommended reconstitution concentrations range from 0.1 mg/mL to 1 mg/mL (100–1000 μg/mL). A commonly used starting point is ≥100 μg/mL.
  • Add the appropriate volume of water gently down the side of the vial. Do not vortex or shake vigorously, as this can denature the protein and reduce activity.
  • Incubate the vial at room temperature for at least 20 minutes to ensure complete dissolution.
  • Mix gently by pipetting up and down or by gentle swirling.

• Aliquoting and Storage:

  • Once fully dissolved, aliquot the protein to avoid repeated freeze-thaw cycles, which can degrade activity.
  • For short-term storage (up to 1 week), keep at 2–8°C.
  • For long-term storage, freeze aliquots at –20°C or –80°C.
  • If desired, add a carrier protein (e.g., 0.1% BSA, 5–10% FBS, 5% HSA, or 5% trehalose) to stabilize the protein during storage and handling.

• Preparation for Cell Culture:

  • Before adding to cells, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or buffer containing a carrier protein if needed.
  • Filter-sterilize the final working solution if sterility is a concern and the protein is compatible with filtration.

Additional Notes:

• Avoid reconstituting at concentrations above 1 mg/mL to prevent solubility issues.
• If solubility is problematic, incubate the solution overnight at 4°C.
• Confirm protein integrity by SDS-PAGE if required.
• Always consult the specific product datasheet for any unique instructions.

Summary Table: Recombinant Mouse IL-16 Reconstitution

These steps will help ensure maximum activity and stability of recombinant mouse IL-16 for your cell culture experiments.