Recombinant Mouse TECK

Recombinant Mouse TECK (also known as CCL25) is a valuable tool for research applications, particularly in immunology and studies of the intestinal immune system. Here are several reasons why you should consider using recombinant Mouse TECK in your research:

1. Specific Biological Function

• Intestinal Immune Regulation: TECK (CCL25) is expressed by intestinal epithelial cells and plays a critical role in recruiting gut-specific memory/effector T cells. Using recombinant Mouse TECK allows you to study the mechanisms of intestinal immune cell trafficking and segmental specialization of the gut immune system.
• Chemotaxis Assays: Recombinant TECK can be used in chemotaxis assays to investigate the migration of T cells and other immune cells in response to this chemokine, providing insights into immune responses in the gut.

2. High Purity and Consistency

• Recombinant Production: Recombinant proteins are produced using well-defined methods, ensuring high purity and consistency. This is crucial for reproducible experimental results and for minimizing batch-to-batch variability.
• Animal-Free Source: Many recombinant proteins, including TECK, can be produced in animal-free systems, reducing the risk of introducing animal-derived pathogens or xenobiotic rejection into your culture system.

3. Controlled Expression and Purification

• Tagged Proteins: Recombinant TECK can be produced with purification tags (e.g., His-tag, FLAG-tag), making it easier to purify and detect the protein in your experiments.
• Controlled Amounts: The amount of recombinant protein can be precisely controlled, allowing you to titrate the concentration and optimize your experimental conditions.

4. Versatile Applications

• Cell Culture Experiments: Recombinant TECK can be used in cell culture to study the effects of this chemokine on T cell migration, activation, and differentiation.
• In Vivo Studies: Recombinant TECK can be administered to animal models to study its effects on immune cell recruitment and gut inflammation in vivo.
• ELISA and Western Blot: Recombinant TECK can serve as a standard in ELISA and as a positive control in Western blot and immunohistochemistry, ensuring the accuracy and reliability of your assays.

5. Research and Therapeutic Insights

• Disease Models: Using recombinant TECK in mouse models can help you understand the role of this chemokine in various diseases, such as inflammatory bowel disease (IBD) and other gut-related disorders.
• Drug Development: Recombinant TECK can be used to screen for potential therapeutic agents that modulate T cell migration and gut immune responses, aiding in the development of new treatments.

6. Compatibility with Other Techniques

• Combination with Antibodies: Recombinant TECK can be used in conjunction with matched antibody pairs for more comprehensive and accurate detection and quantification of the protein in your samples.
• Multiplex Assays: The high specificity and affinity of recombinant TECK make it suitable for use in multiplex assays, where multiple analytes are measured simultaneously.

Summary

Recombinant Mouse TECK is a powerful and versatile tool for studying the intestinal immune system, T cell migration, and gut-related diseases. Its high purity, consistency, and controlled production make it ideal for a wide range of research applications, from basic science to drug development. Using recombinant TECK can enhance the accuracy, reproducibility, and relevance of your research findings.

You can use recombinant Mouse TECK (CCL25) protein as a standard for quantification or calibration in ELISA assays, provided the recombinant protein is of high purity, accurately quantified, and compatible with your assay system. This is a common practice in quantitative ELISA development, especially when natural protein standards are unavailable.

Key considerations for using recombinant Mouse TECK as an ELISA standard:

• Purity and Quantification: The recombinant protein should be highly purified (typically >95% by SDS-PAGE) and its concentration accurately determined, ideally by absorbance at 280 nm or amino acid analysis.
• Formulation: Carrier-free formulations are preferred for ELISA standards to avoid interference from stabilizers like BSA, unless your assay specifically requires them.
• Standard Curve Preparation: Prepare serial dilutions of the recombinant TECK in the same buffer or diluent used for your samples to ensure matrix compatibility and accurate quantification.
• Validation: Confirm that the recombinant TECK produces a standard curve with appropriate sensitivity and dynamic range for your assay. The curve should be linear within the expected concentration range of your samples.
• Parallelism: Ideally, dose-response curves for recombinant and natural TECK should be parallel, indicating similar immunoreactivity in your ELISA system. This ensures accurate quantification of endogenous TECK in biological samples.

Protocol best practices:

• Always follow the ELISA kit or assay protocol for standard preparation, including recommended diluents and incubation conditions.
• Avoid repeated freeze/thaw cycles of the recombinant protein to maintain its integrity.
• Validate recovery and linearity by spiking known amounts of recombinant TECK into representative sample matrices.

Limitations and caveats:

• Recombinant proteins may differ slightly from native proteins in post-translational modifications, which can affect antibody recognition in some assays. Confirm compatibility with your specific ELISA antibodies.
• Use only recombinant proteins specifically intended for ELISA calibration, not those formulated for bioassays or cell culture unless validated for ELISA use.

In summary, recombinant Mouse TECK is suitable as an ELISA standard if it meets purity, quantification, and compatibility requirements, and is validated within your assay system. Always consult your ELISA protocol and perform appropriate controls to ensure accurate quantification.

Recombinant Mouse TECK (CCL25) has been validated primarily for bioassays measuring chemotactic activity, specifically its ability to chemoattract cells expressing the CCR9 receptor, such as Baf3-hCCR9 cells, in a dose-dependent manner. This is the principal application reported in published research and product validation data.

Key validated applications in published research:

• Bioassay (Chemotaxis):

  • The most direct and widely cited application is the use of recombinant mouse TECK in chemotaxis assays, where its ability to attract CCR9+ cells is quantified.
  • This is typically performed using transwell migration assays or similar in vitro systems.

• Functional studies of immune cell trafficking:

  • TECK/CCL25 is used to study the migration of lymphocytes, especially T cells, in the context of thymus and small intestine immune regulation.
  • These studies often use recombinant TECK to stimulate or modulate cell migration in vitro.

Additional notes:

• Carrier-free preparations are often used to avoid interference in sensitive assays.
• No published validation was found for applications such as ELISA, Western blot, immunohistochemistry, or in vivo studies with recombinant protein, although these may be possible with further optimization.

Summary Table:

If you require information on a specific experimental context (e.g., in vivo, immunohistochemistry), please clarify, as current published research and product documentation primarily support chemotaxis/bioassay applications for recombinant mouse TECK.

To reconstitute and prepare Recombinant Mouse TECK protein (also known as CCL25) for cell culture experiments, follow these general best practices for lyophilized recombinant proteins:

• Reconstitution:

  • Use sterile PBS (phosphate-buffered saline) or sterile water as the solvent, unless the product datasheet specifies a different buffer.
  • Aim for a final concentration of 0.1–1.0 mg/mL (100–1000 μg/mL) for the stock solution, as this range is commonly recommended for recombinant proteins used in cell culture.
  • If the protein is supplied with a carrier (such as BSA), reconstitute directly in the buffer. If carrier-free, consider adding 0.1–1% BSA or HSA to prevent adsorption and loss of activity during storage and handling.
  • Briefly centrifuge the vial before opening to collect all lyophilized material at the bottom.

• Dissolving the Protein:

  • Add the calculated volume of buffer gently to the vial.
  • Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation (e.g., slow rotation or gentle pipetting).
  • Do not vortex or shake vigorously, as this can denature the protein or cause foaming.

• Aliquoting and Storage:

  • Once fully dissolved, aliquot the stock solution into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
  • Store aliquots at –20°C or –70°C for long-term storage. For short-term use (up to 1 month), 2–8°C is acceptable if sterility is maintained.
  • Avoid storing at concentrations above 1 mg/mL unless specifically recommended, as this can promote aggregation.

• Working Solution Preparation:

  • Dilute the stock solution to the desired working concentration using cell culture medium or buffer immediately before use.
  • If the protein is sensitive or prone to adsorption, include 0.1% BSA in the working buffer.

• General Handling Tips:

  • Always use sterile technique to prevent contamination.
  • Consult the specific product datasheet for any unique requirements, such as pH, buffer composition, or presence of reducing agents.

Example Protocol for Recombinant Mouse TECK:

1. Briefly centrifuge the vial.
2. Add sterile PBS to achieve 0.1–1.0 mg/mL (e.g., add 100 μL PBS to 100 μg lyophilized protein for 1 mg/mL).
3. Gently mix and let stand at room temperature for 15–30 minutes.
4. Aliquot and store at –20°C or –70°C.
5. Dilute to working concentration in cell culture medium with 0.1% BSA just before use.

Note:
If the datasheet for your specific TECK protein provides different instructions (e.g., a specific buffer or concentration), always follow those recommendations. The above protocol is based on general best practices for recombinant mouse cytokines and chemokines.

If you need the exact buffer composition or concentration for a specific application (e.g., chemotaxis assay, signaling studies), consult the literature or the manufacturer's technical support for guidance.