Recombinant Human CCL22/MDC Protein

Recombinant Human CCL22/MDC Protein is widely used in research to study immune cell recruitment, immune regulation, and disease mechanisms, particularly in immunology, oncology, and inflammation. Its primary value lies in its ability to mimic the natural chemokine CCL22, enabling controlled in vitro and in vivo experiments to dissect its biological functions and signaling pathways.

Key scientific applications and rationale include:

• Immune Cell Recruitment Assays: CCL22 is a potent chemoattractant for monocytes, dendritic cells, natural killer (NK) cells, and regulatory T cells (Tregs), but not for neutrophils or eosinophils. Using recombinant CCL22 allows you to study chemotaxis, cell migration, and the mechanisms of immune cell trafficking under defined conditions.

• Functional Studies of Immune Modulation: CCL22 is involved in the recruitment of Tregs and Th2 cells, which are critical for immune tolerance, suppression of excessive inflammation, and tumor immune evasion. Recombinant protein enables experiments on how CCL22 modulates immune responses, including T cell-DC cluster formation and the establishment of immune privilege in tissues.

• Cancer and Tumor Microenvironment Research: Elevated CCL22 levels are associated with immune cell infiltration in tumors and can correlate with prognosis in certain cancers. Recombinant CCL22 is used to model these processes, test therapeutic interventions, and understand the role of the CCL22-CCR4 axis in tumor immunity.

• Metabolic and Inflammatory Disease Models: Recent studies show that CCL22 can influence adipose tissue beiging, eosinophil recruitment, and anti-inflammatory macrophage (M2) polarization, impacting metabolic homeostasis and inflammation. Recombinant CCL22 is used to dissect these pathways in animal and cell models.

• Bioassays and ELISA Standards: Recombinant CCL22 is essential as a positive control or standard in chemotaxis assays, cytokine profiling, and ELISA quantification.

• Mechanistic Signaling Studies: CCL22 binding to its receptor CCR4 triggers downstream signaling events (e.g., FAK phosphorylation, NF-κB activation), which can be studied in detail using the recombinant protein in cell-based assays.

In summary, using Recombinant Human CCL22/MDC Protein in your research provides a reliable, consistent, and controllable tool to investigate the diverse roles of CCL22 in immune regulation, disease pathogenesis, and therapeutic targeting. This is especially important for dissecting cell-specific responses, validating mechanistic hypotheses, and developing new immunomodulatory strategies.

Yes, recombinant human CCL22/MDC protein can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose in your specific assay system. Most commercial ELISA kits for human CCL22/MDC use recombinant protein as the standard and demonstrate that the assay can accurately quantify both recombinant and naturally expressed CCL22/MDC.

Key considerations and supporting details:

• Recombinant Standard Use: ELISA kits for human CCL22/MDC typically use E. coli-expressed recombinant human CCL22/MDC as the standard, and these kits have shown that the standard curves generated with recombinant protein are parallel to those obtained with naturally expressed CCL22/MDC. This indicates that recombinant protein is suitable for quantification and calibration in these assays.

• Validation: It is essential that the recombinant protein you use as a standard is of high purity, correctly folded, and biologically active. The standard should be validated in your assay to ensure that it produces a linear, reproducible standard curve within the dynamic range of your ELISA.

• Matrix Effects: When using recombinant protein as a standard, it is important to prepare the standard curve in the same matrix as your samples (e.g., serum, plasma, or cell culture supernatant) to account for potential matrix effects that could influence quantification accuracy.

• No International Standard: There is currently no international standard for CCL22/MDC calibration, so the recombinant protein standard you use should be well-characterized and consistent between experiments.

• Documentation: Always refer to your ELISA kit’s instructions and validation data. Many kits specify that both native and recombinant CCL22/MDC are recognized and quantified equivalently by the assay.

Best Practices:

• Prepare a fresh standard curve for each assay run using serial dilutions of the recombinant protein.
• Validate the linearity and recovery of your assay using the recombinant standard in your sample matrix.
• Store and handle the recombinant protein according to manufacturer or supplier recommendations to maintain stability and activity.

In summary: Recombinant human CCL22/MDC protein is widely accepted and used as a standard for ELISA quantification, but proper validation in your specific assay context is critical for accurate results.

Recombinant Human CCL22/MDC Protein has been validated for a range of applications in published research, primarily in studies of immune cell chemotaxis, functional bioassays, and as a standard in immunoassays.

Validated applications include:

• Bioassays / Functional Assays:
  Widely used to assess chemotactic activity, particularly for recruiting regulatory T cells (Tregs), activated T lymphocytes, NK cells, and eosinophils in both in vitro and in vivo models.
  Examples:

  • Recruitment of Tregs in tumor microenvironments and allotransplantation models.
  • Investigation of CCR4 signaling in T cell leukemia/lymphoma.
  • Promotion of beige adipocyte formation and energy metabolism in adipose tissue.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  Used as a standard for quantifying CCL22/MDC levels in biological samples such as serum and cell culture supernatants.
  Example:

  • Measurement of CCL22/MDC in studies of asthma and inflammatory diseases.

• Western Blot:
  Validated for detection and quantification of CCL22/MDC protein expression in cell and tissue lysates.

• Immunohistochemistry:
  Used to localize CCL22/MDC protein in tissue sections, particularly in studies of immune cell infiltration and tumor microenvironments.

• In Vivo Studies:
  Recombinant CCL22/MDC has been administered to animal models to study its effects on immune cell recruitment, thermogenesis, and disease models such as obesity and transplantation.

Additional research applications:

• Positive control in immunological assays.
• Immunogen for antibody production.
• SDS-PAGE and protein characterization.

Summary Table of Validated Applications

Key research areas:

• Immunology (Treg recruitment, chemotaxis, inflammation)
• Oncology (tumor immune privilege, Treg infiltration)
• Metabolism (adipose tissue beiging, energy expenditure)
• Transplantation (immune tolerance)

These applications are supported by multiple peer-reviewed studies and product validation data.

To reconstitute and prepare Recombinant Human CCL22/MDC Protein for cell culture experiments, centrifuge the vial briefly to collect the lyophilized powder at the bottom, then reconstitute with sterile distilled water or sterile PBS to a concentration typically between 0.1–1.0 mg/mL. Do not vortex the solution; instead, gently mix by pipetting or slow inversion.

Step-by-step protocol:

• Centrifuge the vial before opening to ensure all powder is at the bottom.
• Add sterile distilled water or PBS to achieve the desired concentration (commonly 0.1–1.0 mg/mL; some protocols recommend 25–100 μg/mL for working stocks).
• Gently mix by pipetting up and down or slow inversion. Avoid vigorous mixing or vortexing, which can denature the protein.
• If required for stability, add carrier protein (e.g., 0.1% BSA) to the buffer, especially for low concentrations or repeated freeze-thaw cycles.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage: After reconstitution, store at 2–8°C for up to 1 week, or aliquot and freeze at –20°C to –70°C for longer-term storage. Avoid repeated freeze-thaw cycles.

Preparation for cell culture:

• Dilute the stock solution in your cell culture medium to the desired working concentration, typically in the range of 10–100 ng/mL depending on experimental requirements.
• If using serum-free medium, ensure the buffer used for reconstitution is compatible with your cell culture system.
• Sterile filtration (0.2 μm) may be performed after reconstitution if sterility is critical for your application.

Additional notes:

• Always consult the specific product datasheet for recommended reconstitution volumes and buffer conditions, as these may vary depending on the protein formulation and intended use.
• For sensitive applications, consider including a carrier protein (e.g., BSA) to minimize adsorption losses and stabilize the protein in solution.
• Avoid vortexing and repeated freeze-thaw cycles to preserve protein activity.

This protocol ensures optimal solubility and biological activity of recombinant CCL22/MDC for cell culture experiments.