RANTES also known as Chemokine (C-C motif) ligand 5 (CCL5), a CC-chemokine1 is an important mediators of the immune and inflammatory response.2 It is also a potent chemoattractant for T cells 3 and eosinophils.4 RANTES is involved in many inflammatory diseases, for example, bronchial asthma, delayed-type hypersensitivity reactions, viral infections, arthritis, chronic eosinophilic pneumonia, and idiopathic interstitial pneumonia.5
The predicted molecular weight of Recombinant Human Met-RANTES is Mr 7.8 kDa.
Predicted Molecular Mass
7.8
Formulation
This recombinant protein was lyophilized from a 0.2 μm filtered solution in 35% acetonitrile (CH3CN) and 0.1% trifluoroacetic acid (TFA).
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
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Recombinant Human Met-RANTES is a modified form of the chemokine CCL5/RANTES, engineered with an N-terminal methionine, and is widely used in research for its ability to selectively antagonize chemokine receptors, particularly CCR1 and CCR5. Its unique properties make it valuable for dissecting chemokine-mediated inflammatory pathways and for therapeutic modeling in various disease contexts.
Key scientific reasons to use Recombinant Human Met-RANTES:
Selective Chemokine Receptor Antagonism: Met-RANTES acts as a functional antagonist for CCR1 and CCR5, blocking their activation by endogenous ligands. This allows researchers to study the roles of these receptors in immune cell recruitment, inflammation, and tissue injury.
Inflammation and Disease Modeling: Met-RANTES has been shown to reduce tissue inflammation and immune cell infiltration in vivo, making it a powerful tool for modeling diseases such as intracerebral hemorrhage, autoimmune disorders, and chronic inflammatory conditions.
Blood–Brain Barrier Protection: In neurological research, Met-RANTES preserves blood–brain barrier integrity and reduces neurobehavioral deficits following injury by inhibiting CCR1/SRC/Rac1 signaling.
Cancer and Tumor Microenvironment Studies: CCL5/RANTES is implicated in tumor progression and immune cell recruitment in cancer. Met-RANTES can be used to block these effects, aiding studies on tumor microenvironment and metastasis.
HIV Research: The N-terminal modification in Met-RANTES enhances its ability to block M-tropic HIV-1 infection of monocytes, making it relevant for virology and immunology research.
Chemoattraction Assays: Met-RANTES is validated for use in chemotaxis assays, allowing quantification of its inhibitory effects on monocyte and leukocyte migration.
Typical research applications include:
In vivo models of inflammation and tissue injury (e.g., murine models of intracerebral hemorrhage, neuropathic pain, cancer).
In vitro cell migration and chemotaxis assays to study immune cell recruitment.
Mechanistic studies of chemokine receptor signaling and downstream pathways (e.g., SRC/Rac1).
Investigation of chemokine involvement in autoimmune, allergic, and infectious diseases.
Best practices:
Use carrier-free preparations for bioassays or cell culture to avoid interference from stabilizing proteins.
Validate concentration and activity in your specific assay system; typical effective doses range from 0.02–1.2 μg/mL depending on the cell type and assay.
Confirm receptor expression in your model system to ensure specificity of Met-RANTES effects.
Summary: Met-RANTES is a scientifically robust tool for selectively blocking CCR1/CCR5-mediated chemokine signaling, enabling precise investigation of inflammatory, neurological, and oncological processes in both in vitro and in vivo research settings.
Recombinant Human Met-RANTES can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay's antibody specificity and calibration requirements.
Key considerations:
Antibody Specificity: ELISA kits for RANTES (CCL5) typically use antibodies that recognize both natural and recombinant forms, including Met-RANTES, as long as the epitope is preserved. However, Met-RANTES has an N-terminal methionine, which may result in a slight mass difference (+131.2 Da) compared to native RANTES. Most commercial ELISA kits are validated to detect recombinant RANTES, but you should confirm that your kit's antibodies do not discriminate between native and Met-RANTES.
Calibration and Standard Curve: Recombinant proteins, including Met-RANTES, are routinely used to generate standard curves for quantitative ELISA assays. The concentration range and linearity should be validated for your specific assay. Some kits calibrate their standards to international reference preparations (e.g., NIBSC 95/520), so if absolute quantification is required, ensure your recombinant Met-RANTES is traceable or comparable to these standards.
Validation: It is best practice to verify recovery, linearity, and parallelism when using Met-RANTES as a standard in your ELISA system. This ensures that the recombinant standard behaves similarly to endogenous RANTES in your sample matrix.
Documentation: Some ELISA kit protocols explicitly mention the use of recombinant RANTES as a standard and provide guidance on preparation and calibration. Always consult your kit's documentation for compatibility and recommended procedures.
Summary of best practices:
Confirm antibody cross-reactivity with Met-RANTES.
Validate standard curve performance (linearity, recovery, parallelism).
If using for clinical or regulatory purposes, ensure traceability to recognized reference standards.
In conclusion: Recombinant Human Met-RANTES is suitable as a standard for ELISA quantification, but assay-specific validation is essential for accurate and reproducible results.
Recombinant Human Met-RANTES (CCL5) has been validated for a range of applications in published research, primarily focusing on its role as a chemokine modulator and its effects in inflammatory and disease models. Key applications include:
In Vivo Studies: Met-RANTES has been used in murine models to investigate its effects on immune-mediated tissue inflammation, neuropathic pain, and intracerebral hemorrhage (ICH). It has demonstrated efficacy in reducing brain edema, preserving blood-brain barrier (BBB) integrity, and improving neurological outcomes in ICH models. It has also been shown to attenuate inflammatory and nociceptive responses in neuropathic pain models.
Bioassays: The protein is validated for use in bioassays, such as chemotaxis assays, where it has been shown to act as a partial agonist in monocyte chemotactic assays. It has also been used to study the effects of CCL5 on cell migration and activation in various cell types, including cancer-associated fibroblasts and immune cells.
Cell Culture: Met-RANTES is used in cell or tissue culture to study its effects on cell signaling, migration, and activation. It is often recommended with BSA for these applications to maintain protein stability.
ELISA Standards: The recombinant protein is used as a standard in ELISA assays to quantify CCL5 levels in biological samples.
Mechanistic Studies: Research has utilized Met-RANTES to explore the signaling pathways involved in inflammation and disease progression, such as the CCR1/SRC/Rac1 pathway in BBB integrity and the role of CCL5 in cancer progression and cisplatin resistance.
These applications highlight the versatility of Recombinant Human Met-RANTES in both basic research and preclinical studies, particularly in the fields of immunology, neuroscience, and oncology.
To reconstitute and prepare Recombinant Human Met-RANTES protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA). This stabilizes the protein and minimizes adsorption to plastic surfaces.
Step-by-step protocol:
Equilibrate: Allow the vial and PBS (with 0.1% BSA) to reach room temperature before opening.
Centrifuge: Briefly centrifuge the vial (3000–3500 rpm, 5 min) to collect all lyophilized material at the bottom.
Add buffer: Add the calculated volume of sterile PBS + 0.1% BSA to achieve 100 μg/mL. For example, add 100 μL to 100 μg of protein.
Dissolve: Gently swirl or pipette up and down to mix. Do not vortex, as this can denature the protein.
Incubate: Let the vial stand at room temperature for 15–30 minutes with gentle agitation to ensure complete dissolution.
Aliquot: Divide into single-use aliquots (≥20 μL) to avoid repeated freeze-thaw cycles.
Storage: Store aliquots at −20 °C to −70 °C for long-term use, or at 2–8 °C for up to one month. Avoid repeated freeze-thaw cycles.
Dilution for cell culture:
After reconstitution, further dilute the protein in cell culture medium or PBS containing 0.1% BSA to the desired working concentration (e.g., 0.02–1.2 μg/mL, depending on your assay).
For serum-free applications, use a non-protein stabilizer such as trehalose instead of BSA.
Additional notes:
If the protein does not fully dissolve, gently mix for a longer period at room temperature, then at 4 °C overnight if needed.
Avoid vigorous shaking or foaming, which can denature the protein.
For long-term storage, consider adding 5–50% glycerol as a cryoprotectant.
This protocol ensures optimal solubility, stability, and biological activity of Met-RANTES for cell culture experiments.
References & Citations
1. Lee, SC. et al. (2004) J Neurochem.90: 297 2. Borkowski, J. et al. (2004) Arch Immunol Ther Exp (Warsz)52: 201 3. Geiger, H. et al. (2005) Kidney Blood Press Res28: 48 4. Hirata, I. et al.(2009) Dig Dis Sci.54(6):1247-52. 5. Emilie, D. et al. (1994) J Exp Med.179: 1689
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
