Monocyte chemotactic protein-1 (MCP-1) is an essential chemokine produced predominantly by macrophages and endothelial cells and is a potent chemotactic factor for monocytes.1,2,3 It can induce the proliferation and activation of killer cells known as CHAK (CC-Chemokine-activated killer). MCP-1 is also a potent activator of human basophils, induces degranulation and the release of histamines. MCP-1 is involved in the pathogenesis of atherosclerosis and myocardial infarction.4
Protein Details
Purity
>97% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<0.01EU/µg as determined by the LAL method
Biological Activity
Measured by its ability to chemoattract BaF3 mouse pro‑B cells transfected with human CCR2A. The ED<sub>50</sub> for this effect is typically 2-10 ng/mL.
The predicted molecular weight of Recombinant Mouse MCP-1 is Mr 8.5 kDa.
Predicted Molecular Mass
8.5
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives.
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 Mouse MCP-1 (also known as CCL2) is widely used in research to study immune cell recruitment, inflammation, and disease mechanisms, particularly those involving monocyte and macrophage migration. It is a key chemokine that regulates the migration and infiltration of monocytes/macrophages and other immune cells to sites of inflammation or injury.
Essential context and supporting details:
Chemotaxis and Immune Cell Recruitment: MCP-1 is a potent chemotactic factor for monocytes, memory T cells, and dendritic cells, making it essential for modeling and dissecting immune cell trafficking in vitro and in vivo. This is particularly relevant in studies of inflammation, tissue injury, and immune responses.
Disease Modeling: Recombinant Mouse MCP-1 is used to mimic or modulate inflammatory responses in mouse models of diseases such as atherosclerosis, cancer, and central nervous system (CNS) injury. For example, MCP-1 is implicated in the recruitment of monocytes to atherosclerotic lesions and tumor microenvironments, influencing disease progression and therapeutic responses.
Functional Studies: Using recombinant MCP-1 allows precise control over concentration and timing, enabling detailed studies of its effects on cell signaling, migration, polarization, and activation. For instance, studies have shown that monomeric MCP-1 is more effective at inducing microglial migration and polarization than dimeric forms, highlighting the importance of protein form in functional assays.
Assay Standardization: Recombinant MCP-1 is commonly used as a standard in ELISA, bioactivity assays, and as a positive control in Western blotting or neutralization experiments, ensuring reproducibility and comparability across experiments.
Therapeutic Target Validation: MCP-1/CCR2 signaling is a therapeutic target in various inflammatory and neoplastic diseases. Recombinant MCP-1 is used to validate inhibitors, antibodies, or gene therapies targeting this pathway in preclinical models.
Additional relevant information:
Species Specificity: Recombinant Mouse MCP-1 is specifically required for mouse models due to species differences in chemokine structure and function, ensuring physiological relevance in murine studies.
Mechanistic Insights: Studies using recombinant MCP-1 have elucidated its role in activating downstream signaling pathways (e.g., STAT3, NF-κB), promoting cytokine production (IL-6, TNF-α), and modulating immune cell phenotypes in disease contexts.
Protocol Flexibility: Recombinant MCP-1 can be used in a variety of applications, including in vitro chemotaxis assays, in vivo injection for recruitment studies, and as a tool for dissecting chemokine-receptor interactions.
In summary, using recombinant Mouse MCP-1 enables controlled, reproducible investigation of monocyte/macrophage biology, immune cell trafficking, and disease mechanisms in mouse models, supporting both basic research and preclinical therapeutic development.
Yes, recombinant mouse MCP-1 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to your assay system. Recombinant MCP-1 is widely used as a standard in commercial mouse MCP-1 ELISA kits, and its performance is well-documented in the literature and kit protocols.
Key considerations and supporting details:
Equivalence to Natural MCP-1: ELISA kits designed for mouse MCP-1 typically use recombinant mouse MCP-1 (often E. coli-expressed) as the standard. These kits demonstrate that the dose-response curves for recombinant and natural MCP-1 are parallel, indicating that recombinant MCP-1 is suitable for quantifying both recombinant and endogenous MCP-1 in samples.
Calibration and Quantification: The standard curve in ELISA is generated using serial dilutions of recombinant mouse MCP-1. The sample concentrations are then interpolated from this curve, allowing for accurate quantification.
Validation: It is important to ensure that the recombinant MCP-1 you use as a standard is of high purity, correctly folded, and biologically active. The standard should be prepared and stored according to best practices (e.g., with carrier protein at 5–10 mg/mL to prevent adsorption and degradation).
Assay Compatibility: The recombinant MCP-1 standard should be compatible with the antibodies used in your ELISA. Most commercial kits are validated for both natural and recombinant MCP-1, but if you are developing your own assay, confirm that your antibodies recognize the recombinant form equivalently.
Research Use Only: Note that most recombinant MCP-1 standards and ELISA kits are for research use only and not for diagnostic purposes.
Best Practices:
Always run a fresh standard curve with each assay to account for potential variability.
Store recombinant MCP-1 aliquots at recommended temperatures and avoid repeated freeze-thaw cycles.
If using a recombinant standard from a different source than your kit, verify equivalence by comparing standard curves.
Summary Table: Use of Recombinant Mouse MCP-1 as ELISA Standard
Aspect
Details
Source
Recombinant mouse MCP-1 (commonly E. coli-expressed)
Application
Standard for ELISA calibration and quantification
Validation
Parallelism with natural MCP-1, recognized by ELISA antibodies
Storage
With carrier protein, at recommended temperatures
Research Use
For research only, not for diagnostics
In conclusion, recombinant mouse MCP-1 is a validated and widely accepted standard for ELISA quantification, provided it is compatible with your assay system and handled according to best practices.
Recombinant Mouse MCP-1 (CCL2) has been validated for a range of applications in published research, primarily focused on its role as a chemokine in immune cell recruitment and inflammation. The most commonly validated applications include:
Bioassays: Used to assess chemotactic activity, particularly the recruitment of monocytes, macrophages, and other immune cells. Numerous studies have used recombinant mouse MCP-1 to stimulate cells in vitro or in vivo to study migration, signaling, and functional responses.
ELISA Standard: Employed as a quantitative standard in enzyme-linked immunosorbent assays to measure MCP-1 levels in biological samples.
Western Blot Control: Used as a positive control for detecting MCP-1 in Western blotting applications.
Neutralization Assays: Applied in experiments to neutralize MCP-1 activity and study its functional role in various biological processes.
In Vivo Functional Studies: Administered to mice to investigate its effects on immune cell recruitment, inflammation, wound healing, tumor progression, and disease models such as atherosclerosis, pulmonary fibrosis, and neuropathic pain.
Published research examples validating these applications:
Chemotaxis/Bioassay: Recombinant mouse MCP-1 has been used to chemoattract BaF3 mouse pro-B cells transfected with human CCR2A, and to stimulate migration of primary mouse immune cells in vitro.
In Vivo Studies: Used to modulate immune responses in mouse models, such as promoting macrophage recruitment in wound healing, studying tumor-associated macrophage infiltration, and investigating roles in metabolic and inflammatory diseases.
Disease Models: Validated in studies of atherosclerosis, rheumatoid arthritis, psoriasis, pulmonary fibrosis, and neuroinflammation, where MCP-1’s role in monocyte/macrophage recruitment is central.
ELISA and Western Blot: Used as a standard or control protein for quantifying or detecting MCP-1 in various sample types.
Inflammation and Disease: Models of atherosclerosis, rheumatoid arthritis, psoriasis, pulmonary fibrosis, neuroinflammation, and cancer.
Wound Healing: Restoration of macrophage response and tissue repair.
Biomarker Studies: Used as a standard or control in biomarker quantification assays.
These applications are well-supported by both product validation data and a broad range of peer-reviewed publications.
To reconstitute and prepare Recombinant Mouse MCP-1 (CCL2) protein for cell culture experiments, follow these best-practice steps:
1. Reconstitution of Lyophilized Protein
Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
Add sterile distilled water or PBS:
For most applications, reconstitute to a concentration of 0.1–0.5 mg/mL in sterile distilled water.
Some protocols recommend reconstituting at 100 μg/mL in sterile PBS or PBS containing at least 0.1% BSA (bovine serum albumin) to stabilize the protein and prevent adsorption to plastic.
Gently mix (do not vortex or pipette vigorously) to dissolve the protein. The protein may appear as a thin film; allow it to fully dissolve by gentle swirling or tapping.
Let the solution sit for a few minutes at room temperature to ensure complete dissolution.
2. Aliquoting and Storage
Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
Storage conditions:
Short-term (up to 1 month): 2–8 °C under sterile conditions.
Long-term (up to 3 months): –20 °C to –70 °C.
Avoid frost-free freezers and repeated freeze-thaw cycles.
3. Preparation for Cell Culture
Dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or PBS with 0.1% BSA immediately before use.
Typical working concentrations for cell stimulation or chemotaxis assays are in the range of 1–100 ng/mL, depending on the cell type and experimental design.
Filter sterilize the final working solution if sterility is a concern and the protein is not already in a sterile buffer.
4. Additional Notes
If using for ELISA or as a standard, follow the specific dilution instructions provided in the assay protocol.
For in vivo or sensitive cell culture applications, confirm that the endotoxin level is suitable for your experiment (typically <1 EU/μg).
Summary Table: Key Steps for Recombinant Mouse MCP-1 Reconstitution
Step
Details
Reconstitution
0.1–0.5 mg/mL in sterile water or PBS (with 0.1% BSA if needed)
Mixing
Gentle swirling/tapping; avoid vortexing
Aliquoting
Small volumes to avoid freeze-thaw cycles
Storage
2–8 °C (short-term), –20 °C to –70 °C (long-term)
Working dilution
1–100 ng/mL in cell culture medium or PBS with 0.1% BSA
Sterility
Filter sterilize if necessary
Always consult the specific product datasheet for any unique recommendations regarding your recombinant MCP-1 preparation.
References & Citations
1. Yoshimura, T. et al. (1989) J. Exp. Med.169: 1449
2. Oppenheim, JJ. et al. (1989) J. Exp. Med.169: 1485
3. Rollins, BJ. et al. (1997) Blood90: 909
4. Benjamin, EJ. et al. (2005) Ciculation112: 1113
5. Matsushima K. et al. (1989) J. Exp. Med. 169:1485
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
