Recombinant Mouse IP-10 (CXCL10)

Using Recombinant Mouse IP-10 (CXCL10) in research applications is valuable for studying immune cell recruitment, inflammation, tumor biology, and infectious disease mechanisms in mouse models. CXCL10 is a chemokine that plays a central role in immune cell trafficking and modulation of the tumor microenvironment.

Key scientific reasons to use recombinant mouse CXCL10 (IP-10):

• Immune Cell Chemotaxis: CXCL10 is a potent chemoattractant for CD4+ Th1 cells, CD8+ T cells, NK cells, and NKT cells via the CXCR3 receptor, making it essential for dissecting immune cell migration and infiltration in vitro and in vivo.
• Tumor Immunology: CXCL10 inhibits tumor neovascularization (angiogenesis) and has direct anti-proliferative and angiostatic effects on endothelial cells, contributing to its antitumor properties. Recombinant CXCL10 can be used to study these effects or to enhance immune cell infiltration in tumor models, especially in combination with immune checkpoint blockade therapies.
• Inflammation and Autoimmunity: CXCL10 is upregulated in various inflammatory and autoimmune diseases (e.g., arthritis, type I diabetes, experimental autoimmune encephalomyelitis), making it a useful tool for modeling and understanding these conditions in mice.
• Infectious Disease Research: CXCL10 is a biomarker and effector in infectious diseases, modulating immune responses and pathogen control. Recombinant protein can be used to probe its role in host-pathogen interactions or as a standard in ELISA assays.
• Functional Assays: Recombinant CXCL10 is validated for use in chemotaxis assays, bioassays, and as a standard in ELISA, enabling quantitative and mechanistic studies of chemokine signaling and immune cell function.
• Mechanistic Studies: Recombinant protein allows for controlled, reproducible experiments to dissect CXCL10’s receptor-dependent and -independent effects, including its interaction with glycosaminoglycans and its role in modulating cytokine production by monocytes.

Additional considerations:

• Recombinant CXCL10 is typically endotoxin-tested and formulated for bioactivity, ensuring experimental reproducibility.
• It is suitable for both in vitro and in vivo applications, including cell migration, signaling, and tumor microenvironment modulation.

In summary, recombinant mouse CXCL10 (IP-10) is a versatile reagent for immunology, oncology, and infectious disease research, enabling precise manipulation and analysis of chemokine-driven processes in mouse models.

Yes, recombinant mouse IP-10 (CXCL10) is specifically designed and validated for use as a standard for quantification and calibration in ELISA assays. This is one of the primary applications of recombinant IP-10 proteins in research.

Suitability as an ELISA Standard

Recombinant mouse IP-10 is highly suitable for ELISA standardization because it has been rigorously validated against both natural and recombinant forms of the protein. Studies have demonstrated that results obtained using natural mouse IP-10 show linear curves that are parallel to standard curves generated using recombinant IP-10 standards, indicating excellent correlation between the two forms. This parallel behavior confirms that the recombinant protein accurately represents the native protein for quantification purposes.

Recommended Formulation

For ELISA standard applications, recombinant IP-10 is typically supplied in a formulation containing bovine serum albumin (BSA). This formulation is specifically recommended for use as an ELISA standard, as it provides stability and consistency during the assay procedure.

Assay Performance Characteristics

The recombinant protein enables excellent assay precision. Typical intra-assay precision (coefficient of variation) ranges from 4-6% across different concentration ranges, while inter-assay precision ranges from 7.7-10.3%. These precision metrics demonstrate that recombinant IP-10 standards produce reliable and reproducible calibration curves.

Detection Range and Sensitivity

When used as a standard, recombinant mouse IP-10 supports quantification across a broad dynamic range. Depending on the specific ELISA kit configuration, detection ranges typically span from approximately 7.8 pg/mL to 2000 pg/mL, with sensitivities as low as 6.5 pg/mL. This wide range accommodates both low and high abundance samples in your experimental design.

Recombinant Mouse IP-10 (CXCL10) has been validated for a range of applications in published research, primarily focused on its chemotactic, immunomodulatory, and angiostatic properties. The most commonly validated applications include:

• Bioassays/Functional Assays:
  Used to assess chemotactic activity, particularly the recruitment of T cells (CD4+, CD8+), NK cells, and monocytes/macrophages via CXCR3 receptor activation. This includes in vitro migration assays and in vivo models of immune cell trafficking.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  Employed as a standard for quantifying CXCL10 levels in biological samples, such as tissue homogenates or serum, to monitor inflammatory responses or disease progression.

• Western Blot:
  Used as a positive control or standard for protein detection and quantification in cell lysates or tissue samples.

• In Vivo Assays:
  Applied in animal models to study the role of CXCL10 in disease processes, such as autoimmune diseases, infectious diseases, cancer, and tissue regeneration. These studies often involve administration of recombinant CXCL10 to assess its effects on immune cell recruitment, inflammation, angiogenesis inhibition, and tumor growth.

• Binding Assays:
  Used to characterize interactions with its receptor (CXCR3) or to study ligand specificity and receptor activation.

Supporting details and examples from published research:

• Chemotaxis and Immune Cell Recruitment:
  Recombinant Mouse CXCL10 induces chemotaxis of CXCR3-expressing cells (e.g., Baf3-mCXCR3 transfectants) in a dose-dependent manner, with ED₅₀ values reported for specific cell lines. It is widely used to demonstrate the migration of T cells, NK cells, and monocytes in both in vitro and in vivo settings.

• Disease Models:
  Validated in models of autoimmune diseases (e.g., lupus, type 1 diabetes, multiple sclerosis), infectious diseases (malaria, bacterial infections), and cancer, where it is used to study its role in inflammation, immune cell infiltration, and angiogenesis.

• Angiogenesis and Tumor Studies:
  Used to demonstrate anti-proliferative and angiostatic effects on endothelial cells, both in vitro and in vivo, and to investigate its anti-tumor properties.

• ELISA and Western Blot Controls:
  Frequently used as a standard or positive control in ELISA and Western blot protocols for quantifying or detecting CXCL10 in biological samples.

• Novel Applications:
  Engineered fusion proteins (e.g., CXCL10-mucin-GPI) have been used to enhance NK cell recruitment and as adjuvants in immunotherapy research.

Summary Table:

These applications are well-supported in the literature and by product validation data from multiple sources. If you require protocols or more detailed examples for a specific application, please specify the intended use.

To reconstitute and prepare Recombinant Mouse IP-10 (CXCL10) protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) to a concentration of 50–100 μg/mL, ideally with 0.1–1% carrier protein such as bovine serum albumin (BSA) or human serum albumin (HSA) to enhance stability and prevent adsorption to surfaces.

Detailed protocol:

• Bring the vial to room temperature before opening to minimize condensation.
• Add sterile PBS (or another suitable buffer) directly to the vial to achieve your desired stock concentration, commonly 100 μg/mL.
• Include 0.1–1% BSA or HSA in the buffer if the protein is carrier-free, as this helps stabilize the protein and prevents loss due to adsorption.
• Gently swirl or invert (do not vortex) to dissolve the protein completely. Allow the solution to sit at room temperature for 5–10 minutes to ensure full reconstitution.
• Aliquot the stock solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage:
  • Short-term (up to 1 week): 2–8 °C.
  • Long-term: –20 °C or below, preferably in aliquots.
• Working solution: Dilute the stock to the desired final concentration in cell culture medium immediately before use. If possible, add a small amount of carrier protein to the working solution as well.

Additional notes:

• Avoid repeated freeze-thaw cycles to maintain protein activity.
• If using for ELISA or bioassays, follow the specific dilution and buffer recommendations for your application.
• For cell culture, ensure all solutions are sterile and endotoxin levels are appropriate for your assay.

This protocol is based on standard practices and manufacturer recommendations for recombinant mouse CXCL10/IP-10 proteins.