Recombinant Mouse TARC

Recombinant Mouse TARC (CCL17) is a chemokine widely used in research to study immune cell migration, T cell biology, and disease mechanisms, particularly in models of inflammation, allergy, cancer, and autoimmunity. Its use enables precise, reproducible investigation of TARC’s biological functions and its therapeutic or diagnostic potential in mouse systems.

Key scientific reasons to use recombinant Mouse TARC in research applications:

• Chemotaxis of T Cells: Recombinant Mouse TARC is a potent chemoattractant for antigen-primed T helper cells and T cell lines, allowing researchers to study T cell recruitment and trafficking in vitro and in vivo. This is critical for dissecting mechanisms of immune cell migration in disease models.

• Modeling Inflammatory and Allergic Responses: TARC is constitutively expressed in thymic dendritic cells and upregulated in various inflammatory conditions. It is pivotal in mediating Th2-dominated inflammation, making it valuable for modeling allergic diseases, such as asthma and atopic dermatitis, as well as chronic inflammatory states.

• Cancer Immunology: In mouse tumor models, recombinant TARC enhances antitumor immunity, especially when combined with other immunomodulators like GM-CSF. It increases survival, promotes tumor rejection, and augments both innate and adaptive immune responses, including CD4+ and CD8+ T cell infiltration into tumors.

• Autoimmunity and Disease Pathogenesis: Elevated TARC levels are associated with autoimmune diseases, and recombinant TARC can be used to probe its role in disease onset and progression, as well as to test therapeutic interventions targeting the CCL17/CCR4 axis.

• Mechanistic Studies: Recombinant TARC enables controlled experiments to elucidate signaling pathways (e.g., MAPK, NF-κB) activated by TARC-CCR4/CCR8 interactions, and to study its effects on cytokine production, T cell polarization, and regulatory T cell function.

• Standardization and Reproducibility: Using recombinant protein ensures batch-to-batch consistency, defined activity, and the ability to titrate doses for quantitative studies, which is essential for reproducible results in chemotaxis assays, in vivo models, and mechanistic studies.

• Versatility in Assays: Recombinant Mouse TARC is suitable for a range of applications, including:

  • Chemotaxis and migration assays
  • In vivo disease modeling (e.g., allergy, cancer, autoimmunity)
  • ELISA standards and controls
  • Functional studies of T cell and dendritic cell biology

In summary, recombinant Mouse TARC is a critical tool for dissecting immune mechanisms, modeling disease, and evaluating therapeutic strategies in mouse systems, due to its well-characterized, reproducible activity and its central role in T cell-mediated immunity.

Yes, recombinant mouse TARC (CCL17) is commonly used as a standard for quantification or calibration in ELISA assays designed to measure mouse TARC. This is standard practice in commercial ELISA kits and is supported by multiple technical documents and protocols.

Key points and supporting details:

• Recombinant mouse TARC is the calibrator in validated ELISA kits: Commercial mouse TARC ELISA kits use highly purified, E. coli-expressed recombinant mouse TARC as the standard for generating the calibration curve. The assay is calibrated against this recombinant protein, and the standard curve is used to quantify TARC in biological samples.

• Parallelism and equivalence to natural TARC: Validation data show that the standard curves generated with recombinant mouse TARC are parallel to those obtained with natural mouse TARC in biological samples, indicating that the recombinant protein is suitable for accurate quantification. This parallelism is essential for reliable quantification.

• Assay specificity: These ELISA kits are designed to recognize both endogenous (natural) and recombinant mouse TARC, ensuring that the standard curve generated with recombinant protein is applicable to the measurement of native TARC in samples.

• Best practices:

  • Use the recombinant TARC standard provided or validated for your specific ELISA kit, as the antibodies and assay conditions are optimized for this protein.
  • Prepare the standard curve according to the kit instructions, typically by serial dilution of the recombinant TARC stock solution.
  • Avoid mixing standards or reagents from different kits or lots, as this can affect assay accuracy.

• Limitations:

  • The recombinant standard is for research use only and not for diagnostic procedures.
  • If using a recombinant TARC standard from a different source than your ELISA kit, ensure it is of comparable purity and biological activity, and verify parallelism with your assay.

Summary Table: Use of Recombinant Mouse TARC as ELISA Standard

In summary: You can use recombinant mouse TARC as a standard for quantification or calibration in your ELISA assays, provided it is validated for your assay system and you follow the recommended protocols.

Recombinant Mouse TARC (CCL17) has been validated for several key applications in published research, primarily involving its role as a chemokine in immune cell migration and function. The most commonly validated applications include:

• Chemotaxis/Bioassay:
  Recombinant Mouse TARC is widely used in chemotaxis assays to demonstrate its ability to attract T cell lines and antigen-primed T helper cells, particularly via CCR4 and CCR8 receptors. For example, it has been used to chemoattract BaF3 mouse pro-B cells transfected with human CCR4 in a dose-dependent manner.

• In Vivo Functional Studies:
  It has been administered in mouse models to study immune cell recruitment, inflammation, and disease modulation. Published studies have used recombinant TARC in vivo to investigate regulatory T cell chemoattraction, allergic airway inflammation, and immune regulation in disease models.

• ELISA (as Standard):
  Recombinant Mouse TARC is validated as a standard in ELISA assays for quantifying CCL17/TARC levels in biological samples.

• Western Blot:
  It has been used as a positive control or standard in Western blotting to detect CCL17/TARC protein expression.

• Other Functional Assays:
  Additional validated applications include use in blocking assays and immunohistochemistry, as indicated by broader protein validation data.

Summary Table of Validated Applications

Key Published Research Examples:

• Bioassay/Chemotaxis:
  Used to study chemotactic activity for T cells and antigen-primed T helper cells.
• In Vivo:
  Applied in mouse models for selective chemoattraction of regulatory T cells and modulation of allergic airway inflammation.
• ELISA/Western Blot:
  Used as a standard or control in immunoassays for CCL17/TARC quantification and detection.

These applications are well-supported by both product validation data and peer-reviewed publications, confirming the utility of recombinant Mouse TARC in immunological research.

To reconstitute and prepare Recombinant Mouse TARC (CCL17) protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer—commonly sterile PBS or distilled water—at a concentration suitable for your application, typically 25–100 μg/mL. For optimal stability and activity, include 0.1% BSA (bovine serum albumin) in the buffer if the protein is not carrier-free.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all material is at the bottom.
• Add sterile buffer: For most applications, reconstitute at 0.1–1.0 mg/mL in sterile distilled water or PBS. If recommended, use PBS containing at least 0.1% BSA to prevent adsorption and loss of protein activity.
• Gently mix: Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation. Avoid vigorous mixing or vortexing, which can denature the protein.
• Aliquot and store: Once fully dissolved, aliquot the solution to avoid repeated freeze-thaw cycles. Store aliquots at −20°C or colder.
• Working solution: Dilute the stock solution to the desired working concentration in cell culture medium immediately before use.

Additional best practices:

• Do not exceed 1 mg/mL during reconstitution to minimize aggregation.
• Avoid surfactants and organic solvents (e.g., SDS, methanol) in your buffer, as these can denature the protein.
• Carrier-free formulations should be reconstituted in buffer without BSA, but BSA can be added if protein loss is a concern.
• Check product datasheet for specific instructions regarding concentration and buffer, as formulations may vary.

Summary Table:

Always consult the specific product datasheet for any unique requirements. This protocol ensures maximum activity and stability of recombinant Mouse TARC for cell culture experiments.