Recombinant Rat GROβ/MIP-2

Recombinant Rat GROβ/MIP-2 (CXCL2) is a potent neutrophil chemoattractant and activator, making it highly valuable for research applications involving inflammation, immune cell recruitment, and wound healing in rat models.

Key scientific applications and rationale:

• Neutrophil Chemotaxis and Activation: GROβ/MIP-2 is a member of the CXC chemokine family and specifically attracts and activates neutrophils, which are critical for studying acute inflammatory responses, infection models, and tissue injury. Its use enables precise control and quantification of neutrophil migration in vitro (e.g., transwell assays) and in vivo.

• Modeling Inflammatory Diseases: Recombinant rat GROβ/MIP-2 is commonly used to induce or modulate inflammation in experimental models, such as acute lung injury, sepsis, and arthritis. It allows researchers to dissect the molecular mechanisms of neutrophil-driven pathology and test anti-inflammatory interventions.

• Bioassays and Functional Studies: The recombinant protein is suitable for bioassays, including chemotaxis, cell activation, and cytokine release studies. Its defined activity and purity facilitate reproducible results in cell-based assays and animal experiments.

• Receptor-Ligand Interaction Studies: GROβ/MIP-2 binds to CXCR2, enabling investigation of chemokine-receptor signaling pathways, receptor antagonism, and downstream effects on immune cell function.

• Wound Healing and Immune Response Research: By recruiting neutrophils and modulating immune responses, GROβ/MIP-2 is useful for studying tissue repair, host defense mechanisms, and the balance between inflammation and resolution.

Technical considerations:

• Recombinant rat GROβ/MIP-2 is typically produced in E. coli as a non-glycosylated polypeptide, ensuring batch-to-batch consistency and high purity for experimental reproducibility.
• It is fully biologically active and can be used at defined concentrations for dose-response studies and mechanistic investigations.

Summary of best practices:

• Use recombinant rat GROβ/MIP-2 for controlled induction of neutrophil migration and activation in rat-based models.
• Employ it in bioassays to study chemokine signaling, immune cell recruitment, and inflammatory disease mechanisms.
• Select appropriate concentrations and assay formats (e.g., transwell migration, in vivo injection) based on experimental goals and published protocols.

In conclusion, recombinant rat GROβ/MIP-2 is an essential tool for dissecting neutrophil biology, inflammatory processes, and chemokine signaling in rat research applications.

Yes, recombinant rat GROβ/MIP-2 (CXCL2) protein can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known.

Supporting details:

• ELISA quantification requires a standard curve generated from known concentrations of the target protein. Recombinant proteins are commonly used as standards when purified native protein is unavailable.
• Protocols for rat MIP-2 ELISA kits specifically describe the use of recombinant rat MIP-2 as the standard for calibration, with reconstitution and dilution instructions provided to generate a standard curve.
• Validation studies confirm that recombinant rat MIP-2 behaves equivalently to native protein in ELISA quantification, allowing accurate interpolation of sample concentrations.
• Best practices:
  • Ensure the recombinant protein matches the native sequence (e.g., 73 amino acids, non-glycosylated, UniProt P30348).
  • Accurately reconstitute and dilute the standard according to the ELISA kit or assay protocol.
  • Confirm the absence of cross-reactivity or interference from related chemokines (e.g., CXCL1, CXCL3), as validated in commercial kits.
  • Use the same buffer or diluent for standards and samples to minimize matrix effects.

Additional considerations:

• Lot-specific instructions: Always follow the reconstitution and dilution instructions provided with your recombinant protein, as concentration and buffer requirements may vary by lot.
• Research use only: Recombinant standards are suitable for research applications, not for diagnostic use unless specifically validated.

Summary:
Recombinant rat GROβ/MIP-2 is scientifically appropriate and widely used as a standard for ELISA quantification of CXCL2/MIP-2, provided it is properly prepared and validated for your assay system.

Recombinant Rat GROβ/MIP-2 (CXCL2) has been validated in published research primarily for its chemotactic activity on rat neutrophils and for use in functional assays, including chemotaxis and cell migration studies.

Key validated applications in published research include:

• Neutrophil Chemotaxis Assays: Recombinant rat GROβ/MIP-2 has been shown to be a potent chemoattractant for rat neutrophils, with activity demonstrated in vitro using concentrations typically in the range of 10–100 ng/mL. This is the most widely validated and cited application in the literature.

• Functional Characterization: The protein has been used to functionally characterize the chemotactic response of neutrophils, confirming its specificity for neutrophil migration and lack of effect on other cell types such as rat alveolar macrophages or human eosinophils.

• ELISA and Western Blot Standards: While not the protein itself, antibodies raised against rat GROβ/MIP-2 have been validated for use in ELISA and Western blot assays, often using the recombinant protein as a standard or positive control.

• Inflammation and Immunology Research: Recombinant rat GROβ/MIP-2 is frequently used in studies investigating the mechanisms of inflammation, neutrophil recruitment, and cytokine signaling in rat models of disease, including acute lung injury, sepsis, and other inflammatory conditions.

Summary Table: Validated Applications

Additional Notes:

• The recombinant protein is not typically validated for diagnostic or therapeutic use; its applications are restricted to basic and preclinical research.
• Published research has not reported chemotactic activity for other cell types such as macrophages or eosinophils, supporting its specificity for neutrophils.

If you require details on a specific disease model or experimental protocol, please specify the context for more targeted information.

To reconstitute and prepare Recombinant Rat GROβ/MIP-2 (CXCL2) for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
2. Reconstitute the protein using sterile distilled water or the buffer recommended in the product datasheet. Most protocols recommend reconstituting to a concentration between 0.1–1.0 mg/mL. For example, add 100–1000 µL of water to 100 µg of protein.
3. Gently mix by swirling or tapping; avoid vigorous shaking or vortexing to prevent foaming and protein denaturation.
4. Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
5. If required for your application, further dilute the reconstituted protein in cell culture medium or buffer containing carrier protein (e.g., 0.1% BSA or HSA) to minimize adsorption and stabilize the protein.
6. Aliquot the solution to avoid repeated freeze-thaw cycles, which can reduce activity.
7. Storage after reconstitution:
   • Short-term (up to 1 month): Store at 2–8 °C under sterile conditions.
   • Long-term: Store aliquots at –20 °C to –80 °C. Addition of carrier protein is recommended for long-term storage to maintain stability.

Additional notes for cell culture use:

• Confirm the endotoxin level is suitable for your application (typically <1 EU/µg for cell culture).
• Prepare working dilutions in cell culture medium immediately before use.
• Avoid multiple freeze-thaw cycles by aliquoting appropriately.

Summary protocol:

1. Briefly centrifuge vial.2. Add sterile distilled water (or recommended buffer) to achieve 0.1–1.0 mg/mL.3. Gently mix and let dissolve at room temperature for 15–30 min.4. Aliquot and store at 2–8 °C (short-term) or –20 °C to –80 °C (long-term, with carrier protein).5. Dilute to working concentration in cell culture medium just before use.

Always consult the specific product datasheet for any unique instructions or buffer requirements, as some preparations may differ.