Recombinant Mouse/Rat RANTES

Recombinant Mouse/Rat RANTES (CCL5) is widely used in research applications due to its critical role as a chemokine in immune cell recruitment, inflammation, tissue regeneration, and disease modeling in rodent systems.

Key scientific reasons to use recombinant Mouse/Rat RANTES:

• Species-specific activity: Using recombinant RANTES from mouse or rat ensures optimal interaction with rodent chemokine receptors (CCR1, CCR3, CCR5), providing physiologically relevant results in mouse or rat models.
• Immune cell recruitment: RANTES is a potent chemoattractant for T cells, eosinophils, monocytes, and other leukocytes, making it essential for studies on inflammation, immune response, and autoimmune diseases in rodents.
• Modeling inflammatory diseases: RANTES is implicated in various rodent models of inflammatory diseases, such as asthma, delayed-type hypersensitivity, and neuroinflammation, allowing researchers to dissect disease mechanisms and test therapeutic interventions.
• Tissue regeneration and angiogenesis: Recombinant RANTES promotes revascularization and muscle regeneration in injured mouse limbs, and has pro-angiogenic effects relevant for studies on tissue repair and cancer biology.
• Neurobiology and CNS injury: RANTES is involved in axonogenesis, neuronal restoration after brain injury, and regulation of blood–brain barrier integrity, making it valuable for neurobiology and CNS disease research.
• Cancer and tumor microenvironment: RANTES modulates immune cell infiltration and tumor growth in rodent cancer models, supporting studies on tumor immunology and metastasis.
• Stem cell recruitment: Local delivery of RANTES in rats promotes targeted recruitment of mesenchymal stem cells (MSCs) to sites of tissue degeneration, useful for regenerative medicine research.
• Bioassays and functional studies: Recombinant RANTES is used in cell-based assays to study chemotaxis, receptor binding, and signaling pathways in mouse or rat cells, ensuring species compatibility and reproducibility.

Best practices:

• Use recombinant mouse/rat RANTES for in vivo and in vitro studies involving rodent cells or animal models to ensure accurate receptor-ligand interactions and biological responses.
• Select recombinant proteins with validated activity and purity for quantitative and mechanistic assays.

Summary:
Using recombinant Mouse/Rat RANTES enables precise modeling of chemokine-driven processes in rodent systems, supporting research in immunology, inflammation, neurobiology, cancer, and regenerative medicine with species-appropriate reagents.

You can use recombinant Mouse/Rat RANTES as a standard for quantification or calibration in ELISA assays, provided that the ELISA is validated for both recombinant and natural forms of RANTES.

The key points supporting this are:

• Parallelism and Quantification: ELISA kits designed for mouse/rat RANTES typically use recombinant RANTES (expressed in E. coli) as the standard. These kits have demonstrated that the standard curves generated with recombinant RANTES are parallel to those obtained with natural RANTES, indicating that the assay can accurately quantify both forms.
• Calibration: The immunoassay is calibrated against a highly purified recombinant mouse RANTES, and the results show that natural and recombinant proteins yield comparable, linear responses in the assay.
• Relative Mass Determination: The kits are validated to determine the relative mass values of natural mouse and rat RANTES using the recombinant standard, which is essential for accurate quantification in research applications.
• Assay Specificity: The assay is specific for mouse and rat RANTES, recognizing both natural and recombinant forms without significant cross-reactivity or interference from other proteins.

Best Practices:

• Always use the recombinant standard provided or recommended by the ELISA kit manufacturer to ensure compatibility with the assay antibodies and buffers.
• Prepare the standard curve according to the kit protocol, typically by serial dilution of the recombinant RANTES standard.
• Confirm that your assay demonstrates parallelism between the standard curve and your sample dilutions, especially if using a recombinant standard not supplied with the kit.

Limitations:

• The recombinant standard should be of high purity and properly quantified.
• Do not mix standards or reagents from different kits or manufacturers, as antibody specificity and assay conditions may vary.

In summary, recombinant Mouse/Rat RANTES is suitable and commonly used as a standard for ELISA quantification, provided the assay is validated for this purpose and proper protocols are followed.

Recombinant Mouse/Rat RANTES (CCL5) has been validated in published research for a range of applications including bioassays, in vivo studies, ELISA quantification, functional assays, and immunological models.

Key validated applications in published research:

• Bioassays: Recombinant RANTES is widely used to study chemotactic activity, immune cell recruitment, and signaling via CC chemokine receptors (CCR1, CCR3, CCR4, CCR5) in cell-based assays.
• In Vivo Studies: It has been administered to mice in models of asthma, brain injury, ischemia therapy, kidney injury, and cancer to assess its therapeutic and immunomodulatory effects.
• ELISA Quantification: Recombinant RANTES serves as a standard in ELISA kits for quantifying endogenous RANTES levels in mouse and rat serum, plasma, and tissue homogenates.
• Functional Assays: Used to validate chemotactic and activation effects on leukocytes (T cells, macrophages, eosinophils, basophils), and to study histamine release and degranulation.
• Binding Assays: Employed to characterize ligand-receptor interactions and specificity, including studies on viral chemokine-binding proteins.
• Neutralization Studies: Used to investigate the effects of blocking RANTES activity in neuronal restoration and immune regulation.
• Immunohistochemistry and Western Blot: Validated for detection and localization of RANTES in tissue samples.

Representative published research examples:

• Asthma Models: Intranasal administration of recombinant RANTES in mice modulates IgE levels and eosinophil populations, demonstrating its role in allergic inflammation and resolution.
• Neurobiology: RANTES is essential for axonogenesis and neuronal restoration after brain injury in mouse models.
• Cancer Immunology: Recombinant RANTES is used to study immune cell recruitment and tumor microenvironment modulation.
• Ischemia Therapy: RANTES-loaded microparticles promote angiogenesis and tissue repair in mouse ischemia models.
• Kidney Injury: Recombinant RANTES is used in studies of acute kidney injury and immune cell-mediated tissue repair.

Additional validated uses include:

• Delayed-type hypersensitivity reactions
• Viral infection models
• Arthritis and chronic inflammatory disease models

In summary, recombinant Mouse/Rat RANTES is a versatile tool validated for diverse immunological, neurobiological, and disease model applications in both in vitro and in vivo research settings.

To reconstitute and prepare Recombinant Mouse/Rat RANTES (CCL5) protein for cell culture experiments, follow these best-practice steps:

• Reconstitution Concentration:
  Reconstitute the lyophilized protein at a concentration of 100 μg/mL in sterile PBS (phosphate-buffered saline). Some protocols recommend using sterile water for initial reconstitution, but PBS is generally preferred for cell culture compatibility.

• Carrier Protein (Optional but Recommended):
  To enhance protein stability and prevent adsorption to tube walls, add 0.1%–1% BSA (bovine serum albumin) or HSA (human serum albumin) to the PBS before reconstitution, especially if you plan to store aliquots or use low concentrations.

• Dissolving the Protein:
  Gently swirl or invert the vial to dissolve the protein. Avoid vigorous shaking or vortexing to prevent foaming and protein denaturation. Allow the solution to sit at room temperature for 15–30 minutes with gentle agitation until fully dissolved.

• Aliquoting and Storage:

  • After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
  • Store aliquots at 4°C for short-term use (up to 2–7 days).
  • For long-term storage, keep at –20°C or below.
  • If no carrier protein was added during reconstitution, avoid storing diluted protein for extended periods.

• Working Solution Preparation:
  Dilute the reconstituted stock to the desired working concentration using cell culture medium or sterile PBS with carrier protein as needed. Filter-sterilize if required for your application (use a 0.2 μm filter).

• General Handling Tips:

  • Always use sterile technique to prevent contamination.
  • Change pipette tips between additions to avoid cross-contamination.
  • Avoid repeated freeze-thaw cycles by aliquoting appropriately.

Example Protocol:

1. Add 100 μL sterile PBS (with 0.1% BSA) to 10 μg lyophilized RANTES to achieve 100 μg/mL.2. Gently swirl and let stand at room temperature for 20 minutes.3. Aliquot and store at –20°C for long-term use.4. For experiments, dilute aliquots to the required concentration in cell culture medium.

Note:

• Always consult the specific product datasheet for any manufacturer-specific recommendations regarding reconstitution and storage.
• If using for sensitive cell types or in serum-free conditions, ensure all reagents are endotoxin-free.

These steps will ensure optimal solubility, stability, and biological activity of recombinant RANTES for cell culture applications.