Recombinant Mouse CCL27

Recombinant Mouse CCL27 is a valuable tool for research applications focused on skin immunology, barrier tissue homeostasis, T cell migration, and inflammatory disease models, due to its specific role as a chemokine that regulates immune cell trafficking and tissue-specific immune responses.

Key scientific reasons to use recombinant Mouse CCL27 in research:

• Modeling Skin and Barrier Tissue Immunity: CCL27 is critical for the establishment and maintenance of resident lymphocytes, especially T cells expressing the CCR10 receptor, in the skin and other barrier tissues. Using recombinant CCL27 allows you to study how this chemokine regulates immune homeostasis, T cell localization, and tissue-specific immune responses.

• Studying T Cell Migration and Homing: CCL27 specifically attracts skin-associated memory T lymphocytes by binding to CCR10, making it essential for experiments investigating T cell chemotaxis, homing, and retention in cutaneous and mucosal environments. Recombinant CCL27 can be used in in vitro chemotaxis assays or in vivo models to assess the migration of CCR10+ T cells.

• Inflammatory and Autoimmune Disease Models: Altered CCL27 expression is implicated in skin inflammatory diseases such as atopic dermatitis and psoriasis. Recombinant CCL27 can be used to model or modulate these conditions in mice, helping to dissect the mechanisms of disease pathogenesis and the role of chemokine-mediated immune cell recruitment.

• Tumor Immunology: CCL27 is involved in the recruitment of T cells to skin tumors, influencing tumor immune surveillance and progression. Recombinant CCL27 can be used to study tumor-immune interactions, immune escape mechanisms, and the potential for chemokine-based immunotherapies.

• Functional Assays: Recombinant CCL27 is suitable for in vitro and in vivo functional assays, such as:

  • Chemotaxis assays to measure migration of CCR10+ cells.
  • Adhesion assays to study interactions between endothelial cells and T cells.
  • In vivo administration to modulate immune cell localization and inflammation in mouse models.

• Mechanistic Studies: By adding or neutralizing recombinant CCL27, researchers can dissect the molecular pathways involved in immune cell trafficking, tissue-specific inflammation, and immune homeostasis.

In summary, recombinant Mouse CCL27 is essential for dissecting the mechanisms of immune cell migration, tissue-specific immunity, and inflammatory responses in mouse models, particularly in the context of skin and barrier tissue research.

Yes, recombinant mouse CCL27 can be used as a standard for quantification and calibration in ELISA assays, provided it meets specific quality and preparation criteria.

Suitability as an ELISA Standard

Recombinant proteins are widely accepted for ELISA standard preparation. When using recombinant mouse CCL27 as a calibration standard, the protein should ideally be highly purified to ensure accurate quantification. If the recombinant protein is not fully purified, it can be semi-purified in the laboratory and its concentration should be determined using high-performance liquid chromatography (HPLC) or another validated quantification method.

Standard Curve Preparation

When preparing your standard curve with recombinant mouse CCL27, follow these key practices:

Serial Dilution Protocol: Perform 2-fold serial dilutions of your stock standard to establish a working range appropriate for your assay. Most commercial mouse CCL27 ELISA assays operate within ranges of approximately 23.4–1500 pg/mL, though this may vary depending on your specific kit or custom setup.

Blank Control: Include a blank control (typically your dilution buffer with no protein) as your zero standard (0 pg/mL) to establish the baseline for your standard curve.

Curve Fitting: Use curve-fitting software with a four-parameter algorithm to generate the most accurate standard curve fit. The coefficient of determination (R²) of your standard curve should be ≥0.95 to ensure reliability.

Critical Considerations

• Fresh preparation: Prepare a new standard curve for each experiment to account for potential variations in reagent performance and environmental conditions.
• Protein concentration verification: Accurately determine the concentration of your recombinant protein stock before dilution to ensure precise standard concentrations throughout your curve.
• Storage conditions: Maintain proper storage of your recombinant standard according to manufacturer specifications to preserve protein integrity and activity.

This approach ensures that your recombinant mouse CCL27 standard will provide reliable quantification of endogenous or recombinant CCL27 in your samples.

Recombinant Mouse CCL27 has been validated in published research primarily for applications related to immune cell chemotaxis, skin inflammation models, and immune homeostasis studies, particularly in the context of skin and mucosal tissues.

Key validated applications include:

• In vivo chemotaxis assays: Recombinant CCL27 has been injected intradermally or trans-epidermally in mice to demonstrate its ability to recruit T cells, especially CCR10^+^ lymphocytes, to the skin and to modulate local immune responses. This is a direct demonstration of its chemotactic function in living animals.

• Disease model modulation: In mouse models of skin inflammation (such as imiquimod-induced psoriasis and atopic dermatitis), exogenous recombinant CCL27 has been used to assess its regulatory effects on immune cell infiltration, cytokine production, and disease severity. For example, injection of recombinant CCL27 reduced skin inflammation in an imiquimod-induced model.

• In vitro chemotaxis assays: Recombinant CCL27 has been used in transwell migration assays to quantify its ability to attract T cells, confirming its chemotactic activity in controlled cell culture systems.

• Adjuvant studies: CCL27 has been explored as a host-derived adjuvant to enhance immune responses in vaccine research, highlighting its potential to bolster both systemic and mucosal immunity.

• Tumor immunology: Recombinant or adenovirally-expressed CCL27 has been used in tumor models to study its role in promoting T cell infiltration and suppressing tumor growth.

• ELISA standard and cell culture: Recombinant CCL27 is commonly used as a standard in ELISA assays and as a supplement in cell culture to study its effects on immune cell behavior.

Summary of validated research applications:

• In vivo T cell recruitment and skin inflammation modulation
• In vitro chemotaxis/migration assays
• Disease model intervention (e.g., atopic dermatitis, psoriasis)
• Immune adjuvant studies
• Tumor immunology (T cell infiltration, tumor suppression)
• ELISA standard and cell culture supplement

These applications are supported by multiple peer-reviewed studies and reviews, confirming the utility of recombinant mouse CCL27 in both basic and translational immunological research.

To reconstitute and prepare Recombinant Mouse CCL27 protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial: Briefly spin the lyophilized protein vial to ensure all material is at the bottom before opening.

2. Reconstitution buffer:

   • Use sterile PBS (phosphate-buffered saline) as the primary reconstitution buffer.
   • Alternatively, sterile distilled water or PBS containing 0.1% BSA (bovine serum albumin) can be used to improve protein stability and prevent adsorption to tube walls.

3. Concentration:

   • A common working concentration is 100 μg/mL.
   • Adjust the volume added based on the amount of protein and desired final concentration (e.g., add 100 μL to 100 μg lyophilized protein for 1 mg/mL, or 1 mL for 0.1 mg/mL).

4. Mixing: Gently pipette up and down or swirl to dissolve. Avoid vigorous vortexing to prevent protein denaturation.

5. Aliquoting and storage:

   • After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
   • Store aliquots at -20°C to -70°C for long-term storage.
   • For short-term use (up to one week), store at 2–8°C.

6. Handling tips:

   • If using for cell culture, ensure the final solution is sterile.
   • If the protein is carrier-free, consider adding 0.1% BSA or HSA to prevent loss due to adsorption, especially at low concentrations.
   • Avoid multiple freeze-thaw cycles, as this can reduce activity.

Summary protocol:

1. Centrifuge vial briefly.2. Add sterile PBS (or PBS + 0.1% BSA) to achieve 100 μg/mL.3. Gently mix until fully dissolved.4. Aliquot and store at -20°C to -70°C.5. Avoid repeated freeze-thaw cycles.

Application note:
For cell culture, dilute the reconstituted stock to the desired working concentration in your culture medium immediately before use. Always check for endotoxin levels if using in sensitive cell assays.

If your specific product datasheet recommends a different buffer or concentration, follow those instructions, as formulations may vary between suppliers.