Recombinant Human CCL27

Recombinant Human CCL27 is used in research applications to study immune cell migration, skin inflammation, wound healing, and immune homeostasis, particularly in barrier tissues such as skin and mucosa.

CCL27 (also known as CTACK) is a chemokine primarily expressed by keratinocytes and upregulated during inflammation or skin injury. Its main function is to attract CCR10-positive memory T lymphocytes to the skin, mediating their homing and localization. This makes recombinant CCL27 highly valuable for:

• Investigating immune cell trafficking: CCL27 is critical for the migration and localization of CCR10+ T cells in skin and other barrier tissues, which is essential for immune surveillance and tissue-specific immunity.
• Modeling skin inflammation and diseases: Elevated CCL27 levels are associated with inflammatory skin conditions such as atopic dermatitis, psoriasis, and mycosis fungoides. Recombinant CCL27 can be used to induce or modulate inflammatory responses in vitro or in vivo, aiding the study of disease mechanisms and therapeutic interventions.
• Wound healing studies: CCL27 promotes the migration of keratinocyte precursors from bone marrow to skin, facilitating tissue repair and regeneration.
• Tumor microenvironment research: The CCR10/CCL27 axis influences tumor cell migration, immune evasion, and metastasis, especially in melanoma and myeloma models.
• Functional assays and bioactivity testing: Recombinant CCL27 is suitable for chemotaxis assays, cell-based assays, and studies of cytokine-receptor interactions, enabling precise mechanistic investigations.

Best practices for using recombinant CCL27 include:

• Employing it in migration and chemotaxis assays to quantify T cell or keratinocyte responses.
• Using it in animal models to study immune cell recruitment, inflammation, or wound healing.
• Applying it in co-culture systems to investigate stromal-immune cell interactions, especially in cancer or inflammatory disease contexts.

In summary, recombinant human CCL27 is a versatile tool for dissecting mechanisms of immune cell migration, skin homeostasis, inflammation, and tumor biology, making it highly relevant for immunology, dermatology, and oncology research.

Yes, recombinant human CCL27 can be used as a standard for quantification or calibration in ELISA assays, provided it is purified and its concentration is accurately known. Most commercial ELISA kits for human CCL27 include a lyophilized recombinant human CCL27 standard specifically for generating a standard curve to quantify endogenous CCL27 in samples.

When using recombinant CCL27 as an ELISA standard:

• Ensure the protein is purified and the concentration is measured precisely (e.g., by amino acid analysis or HPLC).
• Follow the manufacturer’s reconstitution instructions for the lyophilized protein.
• The standard curve should cover the expected concentration range of your samples (typically 0–1000 pg/mL or higher, depending on the assay sensitivity and sample type).
• For best results, use a recombinant CCL27 that matches the immunoreactivity of the endogenous protein detected by your ELISA kit.

Some recombinant CCL27 products are specifically formulated with carrier proteins like BSA for use as ELISA standards, while carrier-free versions are recommended for applications where BSA could interfere. Always check the product datasheet or consult the manufacturer for guidance on suitability for ELISA standard use.

Recombinant Human CCL27 has been validated in published research primarily for applications related to immune cell migration, chemotaxis assays, and studies of skin and barrier tissue immune homeostasis.

Key validated applications include:

• Bioassays (Chemotaxis/Migration Assays):

  • Used to assess the chemotactic activity of CCL27 on CCR10+ T cells and other lymphocyte populations in vitro, demonstrating its role in attracting skin-associated memory T lymphocytes and mediating their migration.
  • Applied in studies investigating the inhibition of chemotaxis mediated by other chemokine receptors, such as CXCR3, using transfected cell lines and primary cells.
  • Utilized in research on the migration of keratinocyte precursors and the cooperation with other chemokines (e.g., CCL17) in recruiting cutaneous lymphocyte antigen (CLA)+ memory T cells to the skin.

• In Vivo Functional Studies:

  • Recombinant CCL27 has been injected into animal models (e.g., trans-epidermal injection in mice) to study its effects on skin inflammation and immune cell localization, particularly in models of psoriasis and wound healing.
  • Used to demonstrate the role of CCL27 in establishing and maintaining resident lymphocyte populations in skin and other barrier tissues, as well as its impact on immune homeostasis and inflammatory responses.

• Disease Model Research:

  • Applied in studies of inflammatory skin diseases such as atopic dermatitis, psoriasis, and mycosis fungoides, where CCL27 levels and activity correlate with disease severity and immune cell infiltration.
  • Used to investigate the mechanisms of lymphocyte homing and retention in cutaneous and mucosal tissues.

• Cellular and Molecular Mechanism Studies:

  • Employed to elucidate the interaction between CCL27 and its receptor CCR10, as well as its binding to glycosaminoglycans and other cell surface determinants.
  • Used in research on the regulation of T cell localization within skin microstructures and the recruitment of CCR10+ T cells to skin-draining lymph nodes.

Summary Table: Applications of Recombinant Human CCL27 in Published Research

These applications are supported by both in vitro and in vivo studies, with recombinant CCL27 being a critical tool for dissecting its role in immune cell trafficking, skin immunity, and inflammatory disease mechanisms.

Reconstitution Protocol

Recombinant Human CCL27/CTACK protein is typically supplied as a lyophilized powder and requires proper reconstitution before use in cell culture experiments. Begin by centrifuging the vial briefly to concentrate the powder at the bottom of the tube. This step ensures you capture all the protein material when adding the reconstitution buffer.

The recommended reconstitution concentration is 100 μg/mL. For the reconstitution buffer, use sterile phosphate-buffered saline (PBS) containing at least 0.1% human or bovine serum albumin (BSA) if you are using the carrier-protein formulation. If you are using a carrier-free version, reconstitute in sterile PBS without additional carrier protein. Alternatively, some protocols recommend sterile distilled water as the reconstitution medium, though this is less common for cell culture applications.

Formulation Considerations

Two main formulations are available for CCL27 protein:

Carrier-Protein Formulation: Contains BSA as a carrier protein, which enhances protein stability, increases shelf-life, and allows storage at more dilute concentrations. This formulation is recommended for cell or tissue culture applications.

Carrier-Free Formulation: Does not contain BSA and is recommended when the presence of carrier protein could interfere with your experimental design, such as in certain binding assays or when BSA might affect downstream applications.

Storage and Stability

After reconstitution, store working aliquots at -20°C to -80°C for extended storage, with -80°C being optimal for long-term preservation. Reconstituted protein solutions remain stable for approximately 1 month at 2-8°C under sterile conditions, or up to 3 months at -20 to -70°C.

Critical handling requirement: Avoid repeated freeze-thaw cycles, as these significantly affect protein stability and can cause pH changes and protein denaturation. Prepare aliquots of appropriate size for your experiments to minimize freeze-thaw events. Prepare working aliquots containing at least 10 μL of protein solution to maintain stability.

The lyophilized protein itself is stable for 12 months from the date of receipt when stored at -20 to -70°C.

Pre-Experiment Preparation

Before using the reconstituted protein in cell culture, allow the solution to reach room temperature if it has been stored frozen. For optimal results in bioassays and cell culture applications, the effective concentration range is typically 0.1-0.4 μg/mL, so you may need to dilute your stock solution appropriately for your specific experimental protocol.