Recombinant Mouse IL-7

Recombinant Mouse IL-7 is a valuable tool for research applications due to its critical role in regulating immune cell development, survival, and function. Here are several key reasons why you should consider using Recombinant Mouse IL-7 in your studies:

1. Essential for T Cell Homeostasis and Survival

Recombinant Mouse IL-7 is crucial for the maintenance of naïve and memory T cells, including CD4+ and CD8+ T cells, γδ T cells, NKT cells, and T regulatory cells. It promotes T cell survival by upregulating anti-apoptotic proteins such as Bcl-2, making it ideal for experiments focused on T cell persistence and longevity in culture or in vivo models.

2. Supports B Cell Development

IL-7 is essential for normal murine B cell development, particularly at the pro-B and pre-B cell stages. Using recombinant IL-7 allows researchers to study B cell differentiation, expansion, and survival in bone marrow cultures or in vivo models of hematopoiesis.

3. Enhances Immune Reconstitution

In lymphopenic conditions, IL-7 drives the proliferation and expansion of T cells, helping to restore immune cell numbers. This makes it useful for studies on immune recovery after chemotherapy, irradiation, or in models of immunodeficiency.

4. Modulates Peripheral T Cell Subsets

Administration of recombinant IL-7 can modulate the composition of peripheral T cell subsets and increase T cell receptor (TCR) repertoire diversity. This is beneficial for research on immune responses, vaccine development, and adoptive cell therapies.

5. Improves CAR T Cell and Tumor Immunotherapy Outcomes

Studies have shown that recombinant IL-7, especially long-acting forms, enhances the expansion, persistence, and cytotoxicity of CAR T cells and other adoptive T cell therapies in mouse tumor models. This can lead to improved anti-tumor efficacy and prolonged survival in preclinical cancer studies.

6. Facilitates Mechanistic Studies

Recombinant Mouse IL-7 enables detailed investigation of IL-7 signaling pathways, including STAT5 phosphorylation, regulation of CBL-B and SMURF2, and interactions with the IL-7 receptor (IL-7Rα/CD127 and γc/CD132). This is important for understanding the molecular mechanisms underlying immune cell regulation.

7. Versatile Applications

Recombinant Mouse IL-7 can be used in a wide range of experimental settings, including:

• In vitro T and B cell cultures
• In vivo mouse models of infection, autoimmunity, cancer, and immunodeficiency
• Studies on thymopoiesis and lymphoid organ development
• Evaluation of immune reconstitution and adoptive cell transfer therapies

8. Well-Characterized and Available from Multiple Suppliers

Recombinant Mouse IL-7 is widely available from reputable suppliers and is typically produced in E. coli, ensuring high purity and consistency for reproducible results across experiments.

In summary, Recombinant Mouse IL-7 is an indispensable reagent for immunology research, offering robust support for T and B cell biology, immune reconstitution, and therapeutic development in mouse models.

Yes, recombinant Mouse IL-7 can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay’s requirements. Most commercial Mouse IL-7 ELISA kits are designed to recognize both recombinant and natural forms of mouse IL-7, and their standard curves are typically generated using recombinant IL-7.

Key considerations for using recombinant Mouse IL-7 as an ELISA standard:

• Assay Compatibility: ELISA kits for mouse IL-7 generally specify that their antibodies recognize both recombinant and natural IL-7, ensuring accurate quantification regardless of the source.
• Standard Curve Generation: The standard curve in quantitative ELISA is commonly constructed using recombinant mouse IL-7, and results for natural samples are validated to be parallel to those obtained with recombinant standards.
• Purity and Activity: The recombinant protein used as a standard should be highly purified and its concentration accurately determined. Carrier-free formulations are preferred for ELISA standards to avoid interference from stabilizing proteins.
• Validation: It is essential to confirm that the recombinant IL-7 standard is suitable for your specific ELISA kit. Some kits provide their own recombinant standard, which is validated for use with their antibodies and buffers. If using a recombinant IL-7 from another source, ensure it is comparable in purity, formulation, and biological activity.
• Matrix Effects: When preparing standards, dilute recombinant IL-7 in the same buffer or matrix as your samples to minimize matrix effects and ensure accurate quantification.

Best Practices:

• Always follow the ELISA kit manufacturer’s instructions regarding standard preparation and validation.
• Generate a fresh standard curve for each assay run to account for potential variability.
• Confirm that the recombinant IL-7 matches the sequence and post-translational modifications (if any) relevant to your assay.

Limitations:

• Recombinant proteins intended for ELISA standard use are not always suitable for bioassays, as their biological activity may not be validated for functional assays.
• Using a recombinant standard from a different species or with significant sequence variation may result in inaccurate quantification.

Summary Table: Recombinant Mouse IL-7 as ELISA Standard

In conclusion, recombinant Mouse IL-7 is widely accepted and used as a standard for ELISA quantification, provided it is validated for your specific assay and prepared according to best practices.

Recombinant Mouse IL-7 has been validated for a wide range of applications in published research, primarily related to lymphocyte biology, immune cell expansion, and functional assays. Key applications supported by the literature include:

• T cell expansion: Used for in vitro expansion of naive T cells, memory T cells, and T cell subsets such as CD4+ and CD8+ T cells, as well as regulatory T cells and NKT cells.
• T cell differentiation: Supports the differentiation of T cells, including invariant natural killer T (iNKT) cells and other lymphoid lineages.
• Lymphocyte survival and homeostasis: Regulates the survival and maintenance of naïve and memory T cells, αβ and γδ T cells, NKT cells, innate lymphoid cells (ILCs), and T regulatory cells.
• B cell development: Essential for normal murine B cell development, particularly in early stages.
• Lymphopoiesis and lymphoid homeostasis: Promotes the differentiation of hematopoietic stem cells into lymphoid precursor cells, leading to the generation of T cells, B cells, and other lymphoid lineages.
• Functional assays: Used in bioassays to study IL-7-mediated signaling pathways, immune cell proliferation, and cytokine responses.
• Immune reconstitution studies: Applied in models of lymphopenia and immune depletion to study immune recovery and T cell reconstitution.
• Cancer immunotherapy research: Investigated for its potential to enhance anti-tumor immune responses, including in syngeneic tumor models.
• Vaccine adjuvant and immunorestorative effects: Studied for its ability to augment immune responses and restore immune function in immunodeficient or aged models.
• Adoptive cell therapy: Used to support the expansion and function of T cells for adoptive transfer protocols.

These applications are supported by studies in primary cell cultures, animal models, and functional assays, demonstrating the broad utility of recombinant mouse IL-7 in immunological research.

To reconstitute and prepare Recombinant Mouse IL-7 protein for cell culture experiments, follow these best-practice steps:

• Warm the lyophilized vial to room temperature before opening to prevent condensation.
• Centrifuge the vial briefly to collect all powder at the bottom before opening.

Reconstitution:

• Use sterile PBS (phosphate-buffered saline) as the solvent. For carrier-free preparations, add a carrier protein such as 0.1–1% BSA (bovine serum albumin) or HSA (human serum albumin) to improve stability and prevent adsorption to plastic.
• Common working concentrations for stock solutions are 50–100 μg/mL.
• Do not exceed 1 mg/mL during reconstitution to avoid solubility issues.
• Gently mix by pipetting; do not vortex, as this may denature the protein.
• Allow the protein to fully dissolve at room temperature for several minutes.

Aliquoting and Storage:

• Aliquot the reconstituted solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at –20°C or –80°C for long-term storage; for short-term use (up to 1 week), store at 2–8°C.
• Avoid repeated freeze-thaw cycles to maintain activity.

Preparation for Cell Culture:

• Before use, dilute the stock solution to the desired working concentration in cell culture medium. Ensure the final buffer composition is compatible with your cells.
• Typical working concentrations for IL-7 in cell culture range from 0.1–10 ng/mL, depending on the assay and cell type. Refer to published protocols or titrate as needed for your specific application.

Summary Table: Recombinant Mouse IL-7 Reconstitution

Key notes:

• Always consult the specific product datasheet for any manufacturer-specific recommendations.
• Use sterile technique throughout to prevent contamination.
• If using for bioassays, confirm activity with a small-scale pilot experiment.

These steps will ensure optimal solubility, stability, and biological activity of recombinant mouse IL-7 for cell culture applications.