Recombinant Mouse TL1A

Recombinant Mouse TL1A is a valuable tool for research applications focused on immune regulation, inflammation, and disease modeling, particularly in contexts involving T cell biology, mucosal immunity, and fibrotic or autoimmune disorders.

Key reasons to use recombinant Mouse TL1A in research include:

• Immune Cell Activation and Costimulation: TL1A is a member of the TNF superfamily that binds to the DR3 receptor on T cells, providing a potent costimulatory signal. This interaction enhances T cell proliferation, increases IL-2 responsiveness, and stimulates the secretion of pro-inflammatory cytokines such as IFN-γ, GM-CSF, and TNF-α. These properties make TL1A essential for dissecting T cell activation pathways and adaptive immune responses.

• Modeling Inflammatory and Autoimmune Diseases: TL1A is implicated in the pathogenesis of Th1-mediated diseases (e.g., Crohn’s disease, rheumatoid arthritis) and is upregulated in inflamed tissues. Its administration in mouse models can recapitulate features of these diseases, allowing for mechanistic studies and therapeutic testing.

• Fibrosis and Tissue Remodeling: Recombinant TL1A can directly induce fibrosis and airway remodeling by acting on stromal cells (fibroblasts and epithelial cells), making it useful for studying fibrotic mechanisms and testing anti-fibrotic interventions.

• Apoptosis and Angiogenesis: TL1A can induce apoptosis in endothelial cells and inhibit angiogenesis, providing a model to study vascular biology and cell death pathways.

• Mucosal Immunity and Barrier Function: TL1A is a regulator of mucosal immunity, influencing the balance between immune activation and tolerance at barrier sites such as the gut and lungs. It is involved in the induction of type 2 innate lymphoid cells and Th9 cells, which are important for mucosal defense and pathology.

• Experimental Versatility: Recombinant Mouse TL1A is suitable for a range of applications, including:

  • In vitro assays (e.g., T cell proliferation, cytokine secretion, apoptosis studies)
  • In vivo disease models (e.g., airway inflammation, graft-versus-host disease, intestinal injury)
  • ELISA and flow cytometry as a standard or functional ligand.

• Pathway Analysis: TL1A activates key signaling pathways such as NF-κB and MAP kinases, making it a useful reagent for dissecting intracellular signaling events downstream of TNF receptor family members.

Summary Table: Key Applications of Recombinant Mouse TL1A

Best Practices:

• Use recombinant TL1A at concentrations and in models validated in the literature for your specific application.
• Confirm biological activity (e.g., DR3 binding, cytokine induction) before use in functional assays.
• Consider the role of decoy receptors (e.g., DcR3) in modulating TL1A activity in your system.

In summary, recombinant Mouse TL1A is a critical reagent for dissecting immune mechanisms, modeling disease, and testing therapeutic strategies in mouse systems, especially where TNF superfamily signaling is relevant.

Yes, recombinant Mouse TL1A can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose. Recombinant TL1A is commonly used as a standard in commercial ELISA kits designed to quantify mouse TL1A in biological samples.

Key considerations and supporting details:

• Intended Use in ELISA: Many ELISA kits for mouse TL1A explicitly use recombinant mouse TL1A as the standard for generating the calibration curve, which is then used to quantify TL1A in unknown samples. These kits typically include a recombinant TL1A standard with a defined concentration range (e.g., 15.6–1000 pg/mL or 3.125–100 pg/mL).

• Validation: The recombinant standard must be validated to ensure it is recognized by the capture and detection antibodies in your specific ELISA format. Commercial kits are validated for both natural and recombinant TL1A, ensuring that the recombinant protein behaves similarly to the endogenous protein in the assay.

• Standard Curve Preparation: The standard curve is generated by serially diluting the recombinant TL1A and measuring the optical density (OD) at 450 nm after the ELISA reaction. The concentration of TL1A in unknown samples is then interpolated from this curve.

• Protein Formulation: When using recombinant TL1A as a standard, ensure it is in a suitable buffer and formulation (e.g., with or without carrier proteins like BSA) to match the matrix of your samples and minimize matrix effects.

• Quality and Purity: Use recombinant TL1A of high purity (typically >95%) and low endotoxin levels to ensure assay accuracy and reproducibility.

Best Practices:

• Confirm that your recombinant TL1A is compatible with the antibodies used in your ELISA.
• Prepare the standard curve in the same buffer or matrix as your samples to control for matrix effects.
• Store and handle the recombinant protein according to manufacturer or supplier recommendations to maintain stability and activity.

Summary Table: Use of Recombinant Mouse TL1A as ELISA Standard

In conclusion, recombinant Mouse TL1A is widely accepted and used as a standard for ELISA quantification, provided it is validated for your specific assay system and handled according to best practices.

Recombinant Mouse TL1A has been validated for several key applications in published research, primarily in the fields of immunology, inflammation, and autoimmune disease models.

Validated Applications:

• Bioassays: TL1A is widely used in functional bioassays to study its ability to activate NF-κB and MAP kinase signaling, induce apoptosis in endothelial cells, and modulate cytokine secretion (e.g., IFN-γ, GM-CSF, IL-2, TNF-α, IL-6).
• Protein-Protein Interaction/Binding Assays: TL1A’s interaction with its receptors (DR3, DcR3) is routinely assessed using binding assays, including ELISA-based formats.
• Cellular Activation and Expansion: TL1A is used to stimulate T cells and splenocytes ex vivo, enhancing proliferation and cytokine production, and to study costimulatory effects on IL-2 responsiveness.
• In Vivo Disease Models: Recombinant TL1A is employed in mouse models to investigate its role in:
  • Inflammatory Bowel Disease (IBD) and Crohn’s Disease: TL1A levels correlate with disease severity and are used to induce or modulate gut inflammation.
  • Rheumatoid Arthritis (RA): TL1A knockout or neutralization ameliorates collagen-induced arthritis, implicating TL1A in joint inflammation and autoimmunity.
  • Graft-versus-Host Disease: TL1A enhances graft-versus-host reactivity in mouse models.
  • Allergic Lung Inflammation: TL1A:DR3 signaling is studied in Th2-mediated allergic responses.
  • Fibrosis: TL1A-DR3 signaling on fibroblasts is linked to intestinal fibrosis, and neutralization reduces colonic fibrosis in vivo.
  • Antiviral Immunity: TL1A is essential for efficient development of antiviral CD4+ and CD8+ T-cell responses.
• Flow Cytometry: Used to detect TL1A expression and receptor binding on various cell types.
• ELISA: Quantification of TL1A levels in biological samples and assessment of binding activity.
• Western Blot: Detection and quantification of TL1A protein in cell lysates.
• SDS-PAGE: Assessment of purity and molecular weight of recombinant TL1A preparations.

Research Contexts:

• Autoimmune Disease Mechanisms: TL1A is implicated in the pathogenesis of RA, IBD, psoriasis, and other autoimmune conditions through its effects on cytokine production and immune cell activation.
• Angiogenesis Inhibition: TL1A acts as an angiogenesis inhibitor in endothelial cell models.
• Cell Death and Survival: TL1A promotes apoptosis in endothelial cells and survival in activated T cells via NF-κB-dependent pathways.

Summary Table of Validated Applications

Key Insights:

• TL1A is a versatile reagent for dissecting immune signaling pathways, especially those involving TNF superfamily members.
• Its use spans both in vitro and in vivo models, with strong validation in autoimmune and inflammatory disease research.
• Functional studies often focus on TL1A’s role in cytokine induction, cell survival/apoptosis, and disease modulation.

If you require protocol details or specific experimental setups for any application, please specify the context or disease model of interest.

To properly reconstitute and prepare Recombinant Mouse TL1A protein for cell culture experiments, follow these general best practices based on manufacturer protocols and scientific recommendations:

1. Reconstitution

• Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
• Equilibrate the vial and reconstitution buffer to room temperature.
• Reconstitute the lyophilized protein in sterile distilled water or sterile PBS (phosphate-buffered saline), typically at a concentration of 0.1–0.5 mg/mL (or as specified in the product datasheet).
• For some products, adding a carrier protein (e.g., 0.1% BSA, 5% HSA, or 5–10% FBS) is recommended to stabilize the protein, especially for long-term storage or sensitive assays.
• Gently mix the solution by swirling or pipetting slowly; avoid vortexing or vigorous agitation to prevent protein denaturation.
• Allow the protein to dissolve for 15–30 minutes at room temperature with gentle agitation. If flakes persist, mix for up to 2 hours.

2. Storage

• Short-term storage: Reconstituted protein can be stored at 4–8°C for 2–7 days.
• Long-term storage: Aliquot the reconstituted protein and store at ≤ –20°C (preferably –80°C) to minimize freeze-thaw cycles. Aliquots are typically stable for 3 months at –20°C.
• Lyophilized protein: Store desiccated at –20°C to –80°C for up to 12 months.

3. Preparation for Cell Culture

• Dilute the reconstituted TL1A protein in cell culture medium immediately before use.
• If the protocol recommends, include a carrier protein (e.g., 0.1% BSA or 5% FBS) in the dilution buffer to maintain protein stability.
• Filter-sterilize the diluted protein solution if necessary (using a 0.22 µm filter).
• Use freshly prepared solutions for optimal activity, especially for sensitive assays.

4. Additional Notes

• Always refer to the product datasheet or certificate of analysis (CoA) for specific instructions, as formulations and recommendations may vary by manufacturer.
• Avoid repeated freeze-thaw cycles of the reconstituted protein.

By following these steps, you can ensure the proper reconstitution and preparation of Recombinant Mouse TL1A protein for reliable and reproducible cell culture experiments.