Recombinant Mouse Lymphotactin

Recombinant Mouse Lymphotactin (XCL1) is a chemokine widely used in research for its potent and selective chemotactic activity on lymphocytes, especially T cells, NK cells, and dendritic cells, making it valuable for immunological, cancer, and vaccine studies.

Key reasons to use recombinant mouse lymphotactin in research applications:

• Chemotactic Activity: XCL1 specifically attracts lymphocytes, NK cells, and mouse splenocytes, enabling precise studies of immune cell migration, trafficking, and recruitment in vitro and in vivo.
• Receptor Specificity: XCL1 acts through the XCR1 receptor, which is highly expressed on cross-presenting dendritic cells (DCs). This makes it a powerful tool for investigating DC biology, antigen presentation, and the induction of cytotoxic T cell responses.
• Vaccine and Immunotherapy Research: Recombinant XCL1 can serve as an adjuvant to enhance vaccine efficacy by recruiting XCR1+ DCs, thereby boosting antigen-specific CD8+ T cell responses and memory formation. This has been demonstrated in tumor models and infectious disease studies.
• Comparative Immunology and Preclinical Models: It is essential for mouse model studies to dissect chemokine function, immune cell interactions, and disease mechanisms, including cancer, autoimmunity, and neuroinflammation.
• Functional Assays: Recombinant XCL1 is used in chemotaxis assays, flow cytometry, ELISA development, and cell signaling studies to quantify and manipulate immune responses.
• Modulation of Immune Responses: Studies show that XCL1 can enhance dendritic cell-mediated immune responses, promote cytotoxic T lymphocyte activation, and modulate inflammatory processes, making it relevant for both basic and translational research.

Additional considerations:

• Purity and Activity: Recombinant preparations are typically highly pure and biologically active, ensuring reproducibility and reliability in experimental protocols.
• Research-Only Use: Recombinant mouse lymphotactin is intended for research use only and not for diagnostic or therapeutic applications in humans.

In summary, recombinant mouse lymphotactin is a critical reagent for dissecting immune mechanisms, developing vaccines, and modeling disease processes where lymphocyte recruitment and dendritic cell function are central.

Yes, recombinant Mouse Lymphotactin (XCL1) can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose. Recombinant XCL1 is commonly used as a standard in commercial ELISA kits designed to quantify mouse Lymphotactin in biological samples such as serum, plasma, and cell culture supernatants.

Key considerations for use as an ELISA standard:

• Specificity: ELISA kits for mouse XCL1 are typically validated to detect both natural and recombinant forms of the protein, ensuring that recombinant XCL1 is suitable for generating standard curves.
• Purity and Activity: The recombinant protein should be of high purity (typically >95%) and confirmed to be biologically active, as this affects the accuracy of quantification.
• Standard Curve Preparation: The standard should be diluted in the same buffer or matrix as your samples (e.g., standard diluent buffer) to minimize matrix effects and ensure accurate calibration.
• Validation: It is essential to validate the standard curve for linearity, accuracy, and precision according to established guidelines (e.g., FDA, ICH M10) to ensure reliable quantification.
• Documentation: Refer to the ELISA kit manual or assay protocol for recommended concentrations and preparation of the recombinant standard, as these are optimized for each assay system.

Scientific best practices:

• Always confirm that the recombinant XCL1 standard matches the sequence and post-translational modifications of the endogenous protein, as differences may affect antibody recognition.
• Include appropriate controls and run the standard curve in parallel with your samples for each assay plate.
• If developing a custom ELISA, validate the recombinant standard for recovery, dilution linearity, and parallelism with endogenous XCL1 in your sample matrix.

Summary: Recombinant Mouse Lymphotactin is widely accepted and validated as a standard for ELISA quantification, provided it is properly prepared and validated within your assay system.

Recombinant Mouse Lymphotactin (XCL1) has been validated in published research for several key applications, primarily related to immunology and cell migration assays.

Key validated applications include:

• Chemotaxis assays: Recombinant mouse lymphotactin is widely used to assess its chemotactic activity for lymphocytes, natural killer (NK) cells, and mouse splenocytes. It is commonly employed in bioassays measuring the migration of Ba/F3 mouse pro-B cells transfected with human XCR1, confirming its functional activity as a chemoattractant.

• In vitro immune modulation studies: It has been used to enhance the immune efficacy of dendritoma (fusion of dendritic cells and tumor cells) in vitro, demonstrating its role in boosting cytotoxic T lymphocyte responses against tumor cells.

• Comparative immunology and preclinical therapeutic evaluations: Recombinant mouse lymphotactin serves as a reagent in mouse model studies to investigate immune cell recruitment, dendritic cell function, and the XCL1-XCR1 axis in immune responses.

• HIV-1 inhibition studies: XCL1 has been validated as a conformation-dependent inhibitor of HIV-1, blocking viral attachment and entry into target cells in vitro. This application is more commonly reported for human XCL1, but recombinant mouse XCL1 has also been used in mechanistic studies.

• Bioactivity assays: Its activity is routinely validated by its ability to chemoattract specific cell types expressing XCR1, confirming its use in functional bioassays.

Additional context:

• XCL1 is an inflammatory chemokine secreted by T cells and NK cells, with a specialized role in dendritic cell-mediated cytotoxic immune responses and regulatory T cell development.
• It is not chemotactic for monocytes or neutrophils, distinguishing it from other chemokines.
• Recombinant mouse lymphotactin is also used in studies of self-tolerance mechanisms and immune cell interactions in the thymus.

In summary, the most common validated applications are chemotaxis/bioactivity assays, immune modulation studies (including anti-tumor immunity), and mechanistic studies of chemokine function in mouse models.

To reconstitute and prepare Recombinant Mouse Lymphotactin (XCL1) protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS containing at least 0.1%–1% carrier protein (such as bovine or human serum albumin, BSA/HSA). This helps stabilize the protein and prevents adsorption to surfaces.

Step-by-step protocol:

• Centrifuge the vial briefly to collect all lyophilized material at the bottom before opening.
• Add sterile PBS (pH 7.2–7.4) to achieve a final concentration of 100 μg/mL. For example, add 100 μL PBS to 100 μg protein.
• Include carrier protein: Add BSA or HSA to a final concentration of 0.1%–1% in the PBS to minimize protein loss and enhance stability.
• Mix gently by pipetting up and down or gentle vortexing. Avoid vigorous agitation to prevent denaturation.
• Allow the protein to dissolve for 5–10 minutes at room temperature. Inspect for complete dissolution.
• Aliquot the solution into sterile tubes to avoid repeated freeze-thaw cycles.
• Store aliquots at −20 °C to −80 °C for long-term use, or at 2–8 °C for short-term use (up to 1 week). Avoid repeated freeze/thaw cycles.

Further dilution for cell culture:

• Dilute the stock solution in cell culture medium or assay buffer immediately before use.
• Ensure the final working concentration matches your experimental requirements (e.g., based on ED₅₀ or literature recommendations).

Additional notes:

• If the product is carrier-free, you may reconstitute in sterile water, but PBS with carrier protein is preferred for cell culture applications to maintain stability.
• Always check for endotoxin levels if using in sensitive cell types; most recombinant XCL1 preparations are tested to be <0.1 EU/μg.
• Confirm the absence of precipitate before use; filter through a 0.22 μm filter if necessary.

Summary of key points:

• Reconstitute at 100 μg/mL in sterile PBS + 0.1%–1% BSA/HSA
• Aliquot and store at −20 °C to −80 °C
• Avoid repeated freeze/thaw cycles
• Dilute in cell culture medium immediately before use

This protocol ensures optimal stability and bioactivity of recombinant mouse lymphotactin for cell culture experiments.