Recombinant Mouse MIP-2

Recombinant Mouse MIP-2 (CXCL2) is widely used in research applications to study neutrophil recruitment, inflammation, and immune cell signaling due to its potent chemotactic and activating effects on neutrophils.

MIP-2 is a key chemokine secreted by monocytes and neutrophils at sites of inflammation, acting primarily through the CXCR2 receptor to direct neutrophil migration. Its biological activity is critical for modeling and dissecting mechanisms of acute and chronic inflammatory responses in murine systems. Here are the main scientific reasons to use recombinant Mouse MIP-2 in research:

• Neutrophil Chemotaxis Assays: MIP-2 is a robust chemoattractant for mouse neutrophils, making it ideal for in vitro and in vivo migration assays to quantify and analyze neutrophil recruitment. For example, recombinant MIP-2 is commonly used in transwell migration assays to assess neutrophil motility and chemotactic responses.

• Modeling Inflammatory Diseases: MIP-2 is upregulated in various models of inflammation, such as pulmonary inflammation, glomerulonephritis, sepsis, and tissue injury. Its administration or neutralization allows researchers to manipulate neutrophil influx and study the pathogenesis and resolution of inflammatory diseases.

• Immune Cell Activation: Beyond chemotaxis, MIP-2 activates neutrophils, leading to the release of inflammatory mediators and contributing to tissue injury or regeneration, depending on context. This makes it valuable for studying immune cell signaling and effector functions.

• Tissue Regeneration and Injury Models: MIP-2 has been implicated in hepatocyte proliferation and tissue regeneration following injury, such as in liver and corneal models. Recombinant MIP-2 can be used to investigate mechanisms of tissue repair and regeneration.

• Bioassay Standardization: Recombinant MIP-2 provides a consistent, defined reagent for standardizing bioassays, ELISA development, and binding studies involving chemokine-receptor interactions.

• Cellular Source Identification: MIP-2 is expressed by macrophages and epidermal Langerhans cells, making it useful for studies on cellular sources of chemokines in immune responses.

In summary, recombinant Mouse MIP-2 is essential for mechanistic studies of neutrophil biology, inflammation, immune signaling, and tissue regeneration in mouse models. Its use enables precise control and reproducibility in experimental systems investigating chemokine-driven processes.

Yes, you can use recombinant mouse MIP-2 as a standard for quantification or calibration in your ELISA assays, provided it is properly validated and matched to your assay system. Recombinant mouse MIP-2 is commonly used as a standard in commercial ELISA kits, and these kits are validated to ensure that the recombinant protein produces a standard curve that is parallel to the response of natural mouse MIP-2 in biological samples.

Key considerations and supporting details:

• Parallelism and Quantification: Commercial ELISA kits for mouse MIP-2 (CXCL2) are typically calibrated using highly purified recombinant mouse MIP-2 expressed in E. coli. These kits have demonstrated that the dose-response curves for recombinant and natural MIP-2 are parallel, indicating that the recombinant protein is suitable for generating standard curves and quantifying endogenous MIP-2 in samples.

• Assay Validation: The immunoassays are validated for specificity, sensitivity, and precision using recombinant mouse MIP-2 as the standard. This ensures accurate quantification across a range of sample types, including serum, plasma, and cell culture supernatants.

• Preparation of Standards: When preparing your own standards, ensure that the recombinant protein is reconstituted and diluted in the same buffer or diluent used in your assay protocol to minimize matrix effects and maintain consistency with the standard curve.

• Matrix Effects: If your samples are in a complex matrix (e.g., serum, plasma), it is important to confirm that the recombinant standard behaves similarly to endogenous MIP-2 in your specific matrix. Most validated kits report parallelism between recombinant and native MIP-2 in various matrices.

• Best Practices: Always run a standard curve with each assay, using serial dilutions of the recombinant protein, and interpolate sample concentrations from this curve. Internal controls and replicates are recommended to ensure assay reliability.

Summary Table: Use of Recombinant Mouse MIP-2 as ELISA Standard

In conclusion, recombinant mouse MIP-2 is widely accepted and validated as a standard for ELISA quantification, provided you follow best practices for assay setup and validation in your specific experimental context.

Recombinant Mouse MIP-2 (CXCL2) has been validated in published research for several key applications, primarily related to its role as a chemokine in immunological and cell biology studies.

Validated Applications:

• Bioassays: Used extensively to study neutrophil chemotaxis, activation, and recruitment in vitro and in vivo. MIP-2 is a potent chemoattractant for neutrophils, and its activity is commonly assessed using transwell migration assays, where recombinant MIP-2 induces neutrophil migration across a membrane.

• ELISA (Standard and Development): Recombinant MIP-2 serves as a standard for quantifying endogenous MIP-2 levels in biological samples and for developing or validating ELISA protocols.

• Binding Assays: Used to characterize interactions with chemokine receptors (e.g., CXCR2) and to study receptor-ligand binding kinetics.

• In Vivo Studies: Applied in animal models to investigate leukocyte recruitment, inflammation, and immune responses, including pulmonary inflammation, glomerulonephritis, septic peritonitis, and tissue regeneration.

• Cell Migration/Motility Assays: Validated as a chemoattractant in in vitro chemotaxis assays, particularly for mouse neutrophils.

Representative Published Research Applications:

• Neutrophil Recruitment: MIP-2 is used to induce and study neutrophil influx in models of pulmonary inflammation, glomerulonephritis, and heart transplantation.

• Inflammation and Immune Modulation: Investigations into acute lung injury, septic peritonitis, and colonic ischemia-reperfusion have utilized recombinant MIP-2 to probe chemokine-dependent leukocyte recruitment and cytokine signaling.

• Tissue Engineering and Regeneration: MIP-2 has been used to study immune privilege and regeneration in cartilage tissue engineering.

Summary Table of Validated Applications

Additional Notes:

• Recombinant Mouse MIP-2 is typically used with whole cells, tissue homogenates, or in complex sample types, depending on the experimental design.
• Its activity is most commonly validated by its ability to induce chemotaxis in neutrophils and other leukocytes, and by serving as a standard in immunoassays.

If you require protocols or specific experimental details for any of these applications, please specify the intended use.

To reconstitute and prepare Recombinant Mouse MIP-2 protein for cell culture experiments, first briefly centrifuge the vial to collect the lyophilized powder at the bottom. Reconstitute the protein using sterile water or sterile PBS, depending on the formulation and downstream application:

• For general use, reconstitute in sterile deionized or distilled water at a concentration of 0.1–1.0 mg/mL.
• If the formulation contains acetonitrile/TFA or requires a carrier, reconstitute at 100 μg/mL in sterile PBS; adding 0.1% BSA or HSA is recommended for long-term stability.

Protocol Steps:

• Gently pipette the solution down the sides of the vial to ensure full recovery of the protein; do not vortex.
• Mix until fully dissolved. The protein may appear as a film initially; continue gentle mixing until clear.
• Prepare aliquots of the reconstituted stock to avoid repeated freeze-thaw cycles.
• Store aliquots at ≤ –20°C for long-term use; short-term storage at 2–8°C is acceptable for up to one month.

Preparation for Cell Culture:

• Dilute the stock solution into your cell culture medium to the desired working concentration (e.g., 100 ng/mL for chemotaxis assays).
• If using for sensitive applications or long-term storage, include a carrier protein (e.g., 0.1% BSA or HSA) to stabilize the protein.
• Filter sterilize if necessary, especially if the reconstitution buffer or handling may introduce contaminants.

Additional Notes:

• Always consult the specific product datasheet for formulation details, as some preparations may contain additives (e.g., acetonitrile, TFA, BSA) that influence the choice of reconstitution buffer.
• Avoid repeated freeze-thaw cycles to preserve protein activity.
• For bioassays, typical working concentrations range from 10–100 ng/mL depending on cell type and experimental design.

This protocol ensures optimal recovery and stability of recombinant mouse MIP-2 for cell culture experiments.