Recombinant Mouse sTNF RII

Recombinant Mouse sTNF RII is used in research applications to study and modulate TNF-α signaling, investigate immune and inflammatory responses, and serve as a tool for cytokine neutralization in mouse models.

Key scientific reasons for its use:

• Species specificity: Mouse sTNF RII is homologous to endogenous mouse proteins, ensuring accurate biological interactions and minimizing cross-species artifacts when working with mouse cells or in vivo mouse models.
• Neutralization of TNF-α: Soluble TNF RII binds membrane-bound TNF-α and Lymphotoxin-α/β, acting as a decoy receptor to inhibit TNF-mediated signaling, which is crucial for studying inflammatory pathways and autoimmune disease mechanisms.
• Immunomodulation: sTNF RII can modulate immune cell activation, apoptosis, and pro-survival responses, making it valuable for dissecting the roles of TNF signaling in immune regulation, sepsis, and chronic inflammation.
• Bioassay and ELISA standard: Recombinant sTNF RII is commonly used as a standard in ELISA assays to quantify soluble TNF receptors or TNF-α activity in biological samples, enabling precise measurement of cytokine dynamics.
• Therapeutic modeling: Recombinant sTNF RII is used to model anti-TNF therapies in preclinical studies, such as rheumatoid arthritis, allowing researchers to evaluate efficacy, mechanism, and safety in mouse models before clinical translation.
• Functional studies: Fusion proteins (e.g., elastin-mimetic ELP-sTNFRII) retain immunomodulatory function, supporting applications in drug delivery and tissue engineering research.

Best practices:

• Use recombinant mouse sTNF RII for mouse-specific studies to ensure biological relevance and avoid species mismatch effects.
• Select carrier-free or BSA-containing formulations based on downstream applications (e.g., cell culture, ELISA, or bioassays), as carrier proteins can affect stability and assay interference.
• Store and handle according to recommended protocols to maintain protein activity and reproducibility.

Applications include:

• Cytokine neutralization in mouse models of inflammation and autoimmunity.
• ELISA standard for quantifying soluble TNF receptors.
• Mechanistic studies of TNF signaling in immune cells.
• Preclinical evaluation of anti-TNF therapeutics.

Using recombinant mouse sTNF RII enables precise, reproducible, and biologically relevant investigation of TNF-mediated processes in mouse-based research systems.

Yes, recombinant mouse sTNF RII can be used as a standard for quantification or calibration in ELISA assays, provided it is appropriately formulated and validated for this purpose.

Key considerations and supporting details:

• Intended Use: Recombinant mouse sTNF RII is commonly used as a standard in ELISA assays to generate a standard curve for quantification of sTNF RII in biological samples. Multiple peer-reviewed studies and ELISA kit protocols specify the use of recombinant sTNF RII as the standard for accurate quantification.

• Formulation: For ELISA, it is generally recommended to use the recombinant protein formulated with a carrier protein such as BSA, as this improves stability and reproducibility when preparing standard curves. Carrier-free formulations are available but are more susceptible to adsorption losses and may require additional precautions.

• Validation: The recombinant standard should be validated to ensure that it is recognized equivalently to the native protein by the antibodies used in your ELISA. Commercial ELISA kits and published protocols confirm that recombinant mouse sTNF RII yields linear, accurate standard curves and is suitable for quantifying both recombinant and natural sTNF RII in samples.

• Protocol: Prepare a serial dilution of the recombinant sTNF RII in the same diluent as your samples to generate a standard curve for quantification. Follow the ELISA kit or assay protocol for reconstitution and dilution instructions to ensure consistency and accuracy.

• Documentation: Always refer to the product datasheet or technical manual for specific instructions regarding reconstitution, storage, and use as an ELISA standard.

Limitations and best practices:

• Ensure the recombinant protein matches the sequence and post-translational modifications (if relevant) of the native analyte in your samples, as differences can affect antibody recognition.
• Use freshly prepared or properly stored aliquots to avoid degradation or loss of activity.
• If using a custom or non-kit ELISA, confirm that your antibodies detect both recombinant and native sTNF RII equivalently.

Summary Table: Use of Recombinant Mouse sTNF RII as ELISA Standard

In conclusion, recombinant mouse sTNF RII is widely accepted and validated as a standard for ELISA quantification, provided it is handled and validated according to best practices and assay requirements.

Recombinant Mouse sTNF RII (soluble TNF receptor II, TNFRSF1B) has been validated for several key applications in published research, primarily in the context of immunology and inflammation.

Validated Applications:

• Bioassay: Used to assess biological activity, such as TNF-α neutralization and downstream signaling effects in cell-based systems.
• ELISA (Standard): Employed as a standard or reagent in enzyme-linked immunosorbent assays to quantify soluble TNF RII or to measure TNF-α inhibition in biological samples.
• In vivo Functional Studies: Applied in animal models to investigate its anti-inflammatory effects, such as reducing monocyte infiltration and modulating cytokine profiles in models of acute inflammation (e.g., thioglycolate-induced peritonitis).

Supporting Details:

• Bioassay: Recombinant Mouse sTNF RII is used to validate its ability to bind TNF-α and inhibit its biological activity, which is critical for studies on immune cell activation, apoptosis, and cytokine signaling.
• ELISA: Multiple studies have used recombinant sTNF RII as a standard in ELISA protocols to profile cytokine/inhibitor levels in mouse models of sepsis and inflammation, enabling quantification of circulating sTNF RII and assessment of its regulatory role.
• In vivo Models: Research has demonstrated its utility in modulating inflammatory responses in mouse models, such as reducing monocyte infiltration and altering cytokine concentrations following inflammatory challenge.

Additional Notes:

• Recombinant Mouse sTNF RII is also referenced in the context of therapeutic protein engineering, where fusion proteins containing the soluble ligand-binding domain of TNF RII are used to neutralize TNF-α in preclinical models of autoimmune disease.
• Its role as a decoy receptor for TNF-α makes it a valuable tool for dissecting TNF-mediated signaling pathways and for evaluating anti-TNF strategies in disease models.

Summary Table:

These applications are well-supported in the literature and are foundational for research into TNF biology, inflammation, and therapeutic intervention strategies.

To reconstitute and prepare Recombinant Mouse sTNF RII protein for cell culture experiments, first determine whether your protein is supplied carrier-free or with a carrier protein, as this affects the reconstitution protocol.

General Reconstitution Steps:

1. Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
2. Carrier-Free Protein:
   • Reconstitute at 100 μg/mL in sterile PBS.
   • If you need to dilute further for cell culture, use sterile PBS or cell culture medium.
   • For long-term storage or to prevent protein loss at low concentrations, dilute with a solution containing a carrier protein (e.g., 0.1% BSA, 10% FBS, or 5% HAS).
3. Protein with Carrier (e.g., BSA):
   • Reconstitute at 10 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
   • Mix gently and allow to dissolve at room temperature for 10–30 minutes.
4. Mix gently—avoid vigorous shaking or foaming, which can denature the protein.
5. Aliquot the solution to avoid repeated freeze-thaw cycles.
6. Storage:
   • Short-term: Store at 2–8°C for up to one week.
   • Long-term: Aliquot and store at –20°C to –80°C.
   • For extended storage, adding 5–50% glycerol can help stabilize the protein.

Preparation for Cell Culture:

• Before adding to cells, dilute the reconstituted protein to the desired working concentration using cell culture medium or PBS supplemented with carrier protein (e.g., 0.1% BSA) to minimize adsorption and loss.
• If performing serum-free culture or in vivo experiments, avoid animal-derived carrier proteins and consider using trehalose as a stabilizer.

Best Practices:

• Always consult the product’s Certificate of Analysis (CoA) for specific instructions, as optimal buffer and concentration may vary by batch or supplier.
• Avoid repeated freeze-thaw cycles to maintain protein activity.
• For bioassays, ensure the final buffer is compatible with your cell culture system and does not contain toxic additives.

Summary Table:

Always use sterile technique throughout preparation to avoid contamination.