Recombinant Mouse MIG

Using recombinant Mouse MIG (CXCL9) in research applications provides a controlled, reproducible source of this chemokine for studying immune cell migration, inflammation, and tumor biology in mouse models. Recombinant protein enables precise experimental design and interpretation, especially in immunology, oncology, and cell signaling studies.

Key reasons to use recombinant Mouse MIG in research:

• Chemoattractant Activity: Mouse MIG (CXCL9) is a chemokine induced by IFN-γ that acts as a potent chemoattractant for activated T-lymphocytes and other immune cells, making it essential for studying immune cell trafficking, inflammation, and immune responses in vitro and in vivo.
• Standardization and Reproducibility: Recombinant MIG provides a consistent, well-characterized protein source, reducing variability compared to native or tissue-derived preparations. This is critical for quantitative assays such as ELISA, bioassays, and Western blot controls.
• Functional Studies: Recombinant MIG is used to dissect the role of the CXCL9/CXCR3 axis in various biological processes, including T cell priming, macrophage migration, and tumor microenvironment modulation. It is also valuable for testing the effects of MIG on cell signaling, chemotaxis, and cytokine production in controlled settings.
• Disease Modeling: In mouse models of infection, autoimmunity, cancer, and tissue injury, recombinant MIG can be used to mimic or modulate endogenous chemokine responses, helping to elucidate mechanisms of disease and therapeutic targets.
• Assay Development: Recombinant MIG serves as a standard or positive control in immunoassays (e.g., ELISA) and functional assays, ensuring assay sensitivity and specificity.

Additional considerations:

• Versatility: Recombinant Mouse MIG can be applied in cell culture, animal models, and biochemical assays, supporting a wide range of experimental approaches.
• Mechanistic Insights: By adding or neutralizing recombinant MIG, researchers can directly test hypotheses about its role in immune regulation, angiogenesis, and tumor progression.

In summary, recombinant Mouse MIG is a critical tool for immunological and biomedical research, enabling precise, reproducible studies of chemokine function and immune cell dynamics in mouse systems.

You can use recombinant mouse MIG (CXCL9) as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity, accurately quantified, and compatible with your assay's antibody pair and matrix conditions.

Key considerations and supporting details:

• Recombinant proteins are commonly used as ELISA standards when purified native protein is unavailable. Many commercial ELISA kits for mouse MIG/CXCL9 use recombinant mouse MIG as the standard, and these kits demonstrate that recombinant and native MIG yield parallel standard curves, indicating equivalency for quantification.
• Purity and quantification: The recombinant protein should be highly purified (typically >95%) and its concentration accurately determined, ideally by absorbance at 280 nm or amino acid analysis. Impurities or inaccurate quantification can introduce error into your standard curve.
• Matrix compatibility: The diluent used for the standard curve should match the sample matrix as closely as possible to minimize matrix effects. Some kits recommend evaluating diluents for complex matrices like serum or plasma.
• Validation: If you are developing your own ELISA or using a non-kit format, validate that your recombinant standard produces a linear, parallel standard curve compared to native MIG in your sample type. This ensures accurate quantification.
• Carrier-free vs. carrier-containing: For ELISA standards, carrier-free recombinant proteins are generally preferred to avoid interference from carrier proteins such as BSA.

Limitations:

• Not all recombinant proteins are validated for use as ELISA standards. Some are intended only for bioassays or functional studies and may not be suitable for calibration unless specifically tested for this purpose.
• If using a recombinant standard from a source not validated for ELISA, you should perform a parallelism test to confirm that the standard curve is linear and parallels the response of endogenous MIG in your samples.

Best practice:

• Use a recombinant mouse MIG protein that is specifically labeled or validated for use as an ELISA standard.
• Prepare the standard curve according to established guidelines, ensuring proper reconstitution, dilution, and storage.
• Validate the standard curve in your specific assay context, especially if using custom or in-house reagents.

In summary, recombinant mouse MIG is suitable as an ELISA standard if it is pure, accurately quantified, and validated for your assay conditions.

Recombinant Mouse MIG (CXCL9) has been validated for several applications in published research, primarily in the context of immunological and inflammatory studies. The main applications include:

• Bioassay: The protein is widely used in bioassays to study chemotaxis, particularly for measuring the migration of immune cells such as T cells, NK cells, and macrophages. For example, it has been used to chemoattract BaF3 mouse pro-B cells transfected with mouse CXCR3, as well as human lymphocytes cultured with IL-2.
• ELISA Standard: Recombinant Mouse MIG is used as a standard in ELISA assays to quantify CXCL9 levels in biological samples.
• Western Blot: The protein serves as a control in Western blot experiments to detect CXCL9 expression.
• Functional Assay: It is used in functional assays to investigate the role of CXCL9 in immune cell activation, migration, and signaling pathways.
• In Vivo Studies: Recombinant Mouse MIG has been applied in mouse models to study its effects on disease progression, such as in models of diabetic neuropathy, apical periodontitis, and tumor suppression.
• Immunohistochemistry (IHC): Antibodies against CXCL9/MIG have been used in IHC to analyze tissue expression patterns, such as in mouse meningeal tissue.

These applications highlight the versatility of Recombinant Mouse MIG in both in vitro and in vivo research settings, particularly in immunology and inflammation studies.

Reconstitution Protocol

Recombinant mouse MIG (CXCL9) is supplied as a lyophilized powder and requires proper reconstitution before use in cell culture experiments. Begin by centrifuging the vial before opening to concentrate the powder at the bottom of the tube.

Equilibrate both the vial and your reconstitution buffer to room temperature before proceeding. For the reconstitution buffer, use sterile PBS containing at least 0.1% human or bovine serum albumin (BSA) if using the standard formulation. Alternatively, sterile distilled water or aqueous buffer containing 0.1% BSA can be used. If you are using a carrier-free formulation, reconstitute in sterile PBS without additional BSA.

Reconstitute the protein to a final concentration of 0.1–1.0 mg/mL, though 0.1–0.5 mg/mL is commonly recommended. Add the appropriate volume of buffer to achieve your desired concentration, then allow the protein to reconstitute for 15–30 minutes with gentle agitation. If the powder does not fully dissolve and flakes remain visible, continue mixing for up to 2 hours at room temperature.

Storage and Stability

Short-term storage (up to one week): Store reconstituted protein at 2–8°C.

Long-term storage: Store at –20°C to –70°C in a manual defrost freezer. For extended storage beyond one month, adding a carrier protein such as BSA is recommended to enhance stability.

Critical handling consideration: Avoid repeated freeze-thaw cycles, as these can compromise protein integrity and activity. If you require multiple aliquots, prepare single-use portions to minimize freeze-thaw exposure.

Application Considerations

The biological activity of recombinant mouse MIG is typically measured at concentrations of 0.1–0.5 µg/mL in chemoattraction assays using cells transfected with CXCR3. For cell culture applications, the standard formulation with BSA is recommended, as the carrier protein enhances stability and allows storage at more dilute concentrations. Use the carrier-free formulation only when BSA presence would interfere with your specific experimental design.