Recombinant Mouse TWEAK

Recombinant Mouse TWEAK (TNFSF12) is a valuable tool for a wide range of research applications due to its well-characterized biological functions and its role in key signaling pathways. Here are several reasons why you should consider using Recombinant Mouse TWEAK in your research:

1. Well-Defined Biological Activity

• TWEAK is a member of the TNF superfamily and is known to regulate multiple cellular processes, including apoptosis, inflammation, angiogenesis, and tissue regeneration.
• It is the ligand for the Fn14/TWEAKR (TNFRSF12A) receptor, and its interaction with this receptor mediates downstream signaling pathways such as NF-κB, JAK/STAT, and MAPK, which are critical in disease models and physiological studies.

2. Versatility in Experimental Models

• Recombinant Mouse TWEAK can be used in various experimental systems, including cell culture, organoid models, and in vivo studies.
• It is suitable for investigating the effects of TWEAK in different cell types, such as endothelial cells, osteoblasts, myocytes, and immune cells.

3. High Purity and Bioactivity

• Commercially available recombinant Mouse TWEAK is typically >90% pure, with low endotoxin levels (<0.1 EU/µg), ensuring minimal interference in sensitive assays.
• Its biological activity is validated through functional assays, such as cell proliferation assays using HUVECs (human umbilical vein endothelial cells), confirming its ability to activate relevant signaling pathways.

4. Applications in Disease Modeling

• TWEAK has been implicated in several disease processes, including cancer, atherosclerosis, muscle atrophy, and kidney disease.
• Using recombinant TWEAK allows researchers to study the molecular mechanisms underlying these conditions and to evaluate potential therapeutic interventions.

5. Support for Comparative Immunology and Preclinical Studies

• Recombinant Mouse TWEAK is essential for comparative immunology research, enabling the study of immune responses across species.
• It is also used in preclinical therapeutic evaluations, helping to bridge the gap between basic research and clinical applications.

6. Compatibility with Multiple Assay Types

• Recombinant Mouse TWEAK can be used in a variety of assays, including:
  • Cell proliferation and viability assays
  • Apoptosis and cell death assays
  • Inflammatory cytokine production assays
  • Western blotting and ELISA
  • Functional studies in organoids and in vivo models

7. Ease of Use and Standardization

• Recombinant proteins provide a standardized and reproducible source of TWEAK, reducing variability between experiments.
• They are available in both soluble and membrane-associated forms, allowing flexibility in experimental design.

8. Relevance to Mouse Models

• Given the extensive use of mouse models in biomedical research, recombinant Mouse TWEAK is particularly useful for studies involving genetically engineered mice, where the effects of TWEAK can be directly assessed in a relevant biological context.

In summary, Recombinant Mouse TWEAK is a powerful reagent for studying the biological functions of TWEAK, its role in disease, and its potential as a therapeutic target. Its high purity, bioactivity, and versatility make it an excellent choice for a wide range of research applications.

Yes, recombinant mouse TWEAK can be used as a standard for quantification and calibration in ELISA assays, though there are important considerations for optimal results.

Suitability as an ELISA Standard

Recombinant mouse TWEAK is appropriate for use as an ELISA standard because it is recognized by the antibody pairs used in sandwich ELISA assays designed to measure both natural and recombinant forms of the protein. The recombinant protein will bind to the capture and detection antibodies in the same manner as endogenous TWEAK, allowing for accurate quantification of your samples.

Quality Requirements

When using recombinant mouse TWEAK as a standard, ensure the protein meets appropriate quality specifications. The recombinant protein should demonstrate high purity (typically greater than 95% as determined by SDS-PAGE) and low endotoxin levels (less than 1 EU/µg as determined by LAL testing). These characteristics are critical for reliable standard curve generation and accurate sample quantification.

Formulation Considerations

The formulation of your recombinant protein matters significantly. For ELISA standard applications, recombinant mouse TWEAK formulated with BSA (bovine serum albumin) is recommended. BSA serves as a carrier protein that stabilizes the recombinant protein and prevents non-specific adsorption to assay surfaces, which is particularly important when preparing serial dilutions for standard curves.

Standard Curve Validation

When preparing your standard curve, avoid vortexing the reconstituted standard, as mechanical force can destabilize the protein. To validate that your standard performs correctly, it is recommended to spike the recombinant protein into your sample matrix and verify that the amount spiked in matches the amount detected in the assay. This spike recovery experiment confirms that nothing in your specific sample matrix interferes with the assay and that your standard curve accurately reflects real-world conditions.

Expected Performance

Properly prepared standard curves using recombinant mouse TWEAK typically achieve sensitivity in the range of 0.3 pg/mL with detection ranges extending to approximately 200 pg/mL, depending on the specific antibody pair and assay optimization used.

Recombinant Mouse TWEAK (TNFSF12) has been validated in published research for applications including cell culture, bioassays, in vivo studies, and topical therapeutic models.

Key validated applications in peer-reviewed studies include:

• Cell Culture: Used to study TWEAK signaling in organoids and cultured cells, including effects on muscle atrophy, osteoblast differentiation, and cytokine production.
• Bioassay: Extensively applied to assess biological activity in whole cell systems, such as:
  • Induction of muscle atrophy and cachexia via EDA2R-NIK signaling.
  • Regulation of JAK2/STAT5 and JAK2/STAT3 pathways in hepatocytes and cardiac myocytes.
  • Modulation of RANTES production, BMP-2-induced differentiation, and RANKL expression in osteoblastic cells.
  • Investigation of immune cell activation and cytokine signaling.
  • Assessment of atherosclerotic lesion progression and diabetic complications.
  • Study of crescent formation in IgA nephropathy.
• In Vivo Studies: Validated for direct administration in mouse models to study:
  • Muscle function and pathology in RNA toxicity and ALS models.
  • Kidney fibrosis and profibrotic signaling.
  • Neurovascular permeability via NF-κB pathway activation.
  • Atopic dermatitis and keratinocyte dysregulation.
• Topical Therapeutic Application: Demonstrated efficacy in accelerating burn wound healing in mice, with mechanistic studies on cytokine induction and extracellular matrix synthesis.
• Proliferation Assays: Shown to promote proliferation of human umbilical vein endothelial cells (HUVEC) in dose-dependent manner.
• Immunological and Preclinical Therapeutic Evaluations: Used as a reagent in comparative immunology and preclinical studies.

Summary Table of Validated Applications

Additional Notes:

• Recombinant Mouse TWEAK is typically used in studies involving its receptor Fn14 (TNFRSF12A), and its effects are often validated by genetic deletion or pharmacological blockade in mouse models.
• The protein is widely used to dissect molecular mechanisms in inflammation, tissue remodeling, fibrosis, and immune regulation.
• Protocols may involve direct protein administration, cell culture supplementation, or topical application depending on the experimental design.

These applications are supported by multiple peer-reviewed publications and are considered standard for recombinant Mouse TWEAK in experimental biology.

Reconstitution Protocol

Recombinant Mouse TWEAK should be reconstituted at a concentration of 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA). The lyophilized protein is typically supplied as a 0.2 μm filtered solution that has been lyophilized from PBS and NaCl with BSA as a carrier protein.

Step-by-Step Reconstitution Procedure

Initial Preparation

Begin by allowing both the vial and reconstitution buffer to equilibrate to room temperature before opening. Centrifuge the vial briefly to collect any powder that may have adhered to the walls.

Reconstitution Process

Add the appropriate volume of sterile PBS containing at least 0.1% BSA to achieve your desired concentration of 100 μg/mL. Mix the solution gently by pipetting only—do not vortex or mix vigorously, as this can denature the protein and reduce its biological activity. Allow the reconstituted protein to incubate for 15-30 minutes with gentle agitation to ensure complete dissolution. If visible flakes remain, continue gentle mixing for up to 2 hours at room temperature.

Storage and Stability

Short-Term Storage

After reconstitution, the protein remains stable for 1 month when stored at 2-8°C under sterile conditions. This temperature range is suitable for experiments planned within this timeframe.

Long-Term Storage

For extended storage, maintain the reconstituted protein at -20 to -70°C under sterile conditions for up to 3 months. Use a manual defrost freezer and avoid repeated freeze-thaw cycles, as these can compromise protein integrity and biological activity.

The lyophilized protein itself, when stored desiccated at -20 to -70°C, remains stable for 6-12 months.

Quality Specifications

The recombinant Mouse TWEAK protein typically exhibits the following characteristics relevant to cell culture work:

• Purity: >90% by SDS-PAGE with silver staining
• Endotoxin Level: <0.10 EU per 1 μg by LAL method
• Molecular Weight: Approximately 17 kDa
• Biological Activity: ED₅₀ of 0.03-0.3 μg/mL in functional assays

Preparation for Cell Culture Applications

When preparing the protein for cell culture experiments, further dilutions should be made using culture medium containing 5% fetal calf serum or an appropriate carrier protein to maintain protein stability and prevent non-specific adsorption to culture vessels. This is particularly important for maintaining consistent biological activity across multiple experimental replicates.

Ensure all reconstitution is performed under aseptic conditions to maintain sterility throughout your experiments.