Recombinant Mouse CCL21

Recombinant Mouse CCL21 is widely used in research applications due to its critical role in immune cell trafficking, tumor immunology, and lymphatic biology. It is especially valuable for studies involving immune cell migration, tumor microenvironment modulation, and immunotherapy enhancement.

Key scientific reasons to use recombinant Mouse CCL21:

• Immune Cell Chemotaxis: CCL21 is a potent chemoattractant for dendritic cells (DCs) and naïve T cells, guiding them to lymphoid tissues and sites of inflammation, which is essential for initiating adaptive immune responses. This property is exploited in assays studying immune cell migration, lymph node homing, and peripheral tolerance.

• Tumor Immunology and Therapy: CCL21 can enhance anti-tumor immune responses by recruiting effector immune cells into the tumor microenvironment. Studies show that CCL21 administration, especially in nanoformulated or sustained-release forms, increases intratumoral immune cell infiltration, prolongs survival, and can lead to complete tumor regression in mouse models. It also synergizes with immune checkpoint inhibitors (e.g., anti-PD-1 antibodies) to improve immunotherapy outcomes.

• Modulation of Tumor Microenvironment: CCL21 influences cytokine profiles within tumors, promoting anti-tumor cytokines and reducing pro-tumor cytokines, thereby enhancing the effectiveness of chemotherapy and immunotherapy.

• Lymphatic Biology and Metastasis: CCL21/CCR7 signaling is crucial for lymphatic metastasis in cancer models and for the development and function of secondary lymphoid organs. It is also involved in lymphoid neogenesis and peripheral tolerance.

• Bioassays and Functional Studies: Recombinant CCL21 is biologically active and suitable for chemotaxis assays, in vivo studies, and functional analyses of immune cell behavior. It is commonly used to study DC and T cell migration, lymphatic vessel docking, and immune cell activation.

Typical research applications include:

• Chemotaxis assays for dendritic cells and T cells
• In vivo tumor models to study immune cell infiltration and anti-tumor immunity
• Combination therapy studies with checkpoint inhibitors or chemotherapy
• Investigations of lymphatic metastasis and lymphoid organ development
• Functional studies of immune cell activation and peripheral tolerance

Best practices:
Use recombinant Mouse CCL21 at concentrations validated for your specific assay (e.g., chemotaxis bioassays), and ensure proper controls for specificity. For in vivo applications, consider delivery methods that optimize tissue retention and sustained release, such as nanoformulations or hydrogels.

In summary, recombinant Mouse CCL21 is a versatile tool for dissecting immune cell dynamics, enhancing anti-tumor responses, and studying lymphatic biology in mouse models. Its use is supported by robust evidence in immunology, oncology, and cell migration research.

Recombinant Mouse CCL21 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known. This is a common practice in both commercial ELISA kits and custom assay development.

Key considerations and supporting details:

• Commercial ELISA kits for mouse CCL21 routinely use recombinant mouse CCL21 as the standard for generating calibration curves, enabling quantification of CCL21 in biological samples such as serum, plasma, and cell culture supernatants.
• Recombinant CCL21 protein is specifically recommended for use as an ELISA standard by manufacturers, especially when formulated with BSA for stability.
• Standard curve preparation: The recombinant protein is reconstituted at a known concentration and serially diluted to generate a standard curve, which is then used to interpolate the concentration of CCL21 in unknown samples.
• Assay compatibility: Ensure that the recombinant CCL21 you use matches the sequence and form recognized by the capture and detection antibodies in your ELISA. Most commercial kits specify the amino acid region (e.g., Ser24-Gly133) and the form of CCL21 used as the standard.
• Validation: If you are developing your own ELISA or using a kit with a different standard, validate that your recombinant CCL21 yields a standard curve with expected sensitivity and linearity in your assay system.

Best practices:

• Use carrier-free, endotoxin-free recombinant protein for highest accuracy.
• Confirm the protein’s concentration by an independent method (e.g., absorbance at 280 nm or BCA assay) if absolute quantification is critical.
• Prepare fresh dilutions for each assay to minimize degradation or adsorption losses.

Summary:
You can use recombinant mouse CCL21 as a standard in ELISA assays, as long as it is compatible with your assay’s antibodies and is accurately quantified. This approach is standard in both commercial and custom ELISA protocols.

Recombinant Mouse CCL21 has been validated for a range of applications in published research, primarily focused on its role as a chemokine in immune cell migration and tumor immunology. The most commonly validated applications include:

• Functional/Bioassays: Used to assess chemotactic activity, especially for dendritic cells (DCs), naïve and activated T cells, and in studies of immune cell migration, activation, and signaling.
• ELISA: Utilized as a standard or analyte in enzyme-linked immunosorbent assays to quantify CCL21 or study its interactions.
• Western Blot: Applied for detection and quantification of CCL21 protein, including studies of its expression and cleavage.
• Immunohistochemistry: Used to localize CCL21 in tissue sections, particularly in studies of lymphoid organs and inflamed tissues.
• Blocking Assays: Employed to investigate the inhibition of CCL21-mediated chemotaxis or signaling, often using antibodies or antagonists.
• In vivo Assays: Validated in animal models for studying immune cell trafficking, tumor growth inhibition, and therapeutic delivery (e.g., intratumoral injection, nanoformulation).

Key research applications and findings:

• Tumor Immunotherapy: Intratumoral administration of recombinant CCL21, including nanoformulated versions, has been shown to inhibit tumor growth, enhance immune cell infiltration, and prolong survival in mouse models of neuroblastoma and other cancers.
• Immune Cell Chemotaxis: CCL21 is widely used to study the migration of DCs and T cells, both in vitro and in vivo, due to its potent chemotactic properties.
• Mechanistic Studies: Investigations into the molecular pathways of immune cell migration, lymphatic transendothelial migration, and chemokine receptor signaling frequently employ recombinant CCL21 in bioassays.
• Angiostasis and Dendritic Cell Function: CCL21 has been validated for studies on its angiostatic effects and its role in facilitating DC function and T cell activation.
• Combination Therapies: CCL21 has been used in combination with other treatments (e.g., radiotherapy, adoptive cell therapy) to enhance anti-tumor immune responses.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed studies and product validation data, confirming the utility of recombinant Mouse CCL21 in immunology, oncology, and cell biology research.

To reconstitute and prepare Recombinant Mouse CCL21 protein for cell culture experiments, dissolve the lyophilized protein at a concentration of 100 μg/mL in sterile phosphate-buffered saline (PBS) containing at least 0.1% human or bovine serum albumin (BSA). This formulation helps stabilize the protein and prevents adsorption to surfaces.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (pH 7.2–7.4) containing 0.1% BSA to achieve the desired concentration (e.g., 100 μg/mL).
• Gently mix by pipetting up and down or swirling; avoid vigorous vortexing to prevent protein denaturation.
• Allow the solution to sit at room temperature for several minutes to ensure complete dissolution.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade chemokines.
• Store aliquots at −20°C or below for long-term use; for short-term use, keep at 4°C.

Additional notes for cell culture:

• If your protocol or cell type is sensitive to carrier proteins, you may reconstitute in sterile PBS alone, but including BSA is recommended for stability.
• For bioassays, typical working concentrations range from 3–200 ng/mL, depending on the cell type and experimental design.
• Always use sterile technique to avoid contamination.

Summary Table:

This protocol ensures optimal solubility and biological activity of recombinant CCL21 for cell culture experiments.