Recombinant Mouse CXCL14

Recombinant Mouse CXCL14 is a valuable tool for research because it enables precise investigation of CXCL14’s roles in immune regulation, tumor suppression, antimicrobial defense, muscle physiology, and neurodevelopment. Using the recombinant protein allows for controlled, reproducible experiments in both in vitro and in vivo systems.

Key scientific applications and rationales include:

• Immune Cell Trafficking and Modulation: CXCL14 regulates the migration of macrophage precursors to skin and mucosal tissues, supporting their development and function. It also acts as a chemoattractant for monocytes and immature dendritic cells, influencing immune cell infiltration and tissue homeostasis.

• Tumor Immunology: CXCL14 expression in tumor models enhances recruitment of CD8⁺ T cells and dendritic cells, restoring antigen presentation and promoting antitumor immunity. Loss of CXCL14 is associated with reduced immune cell infiltration and increased tumor growth, highlighting its potential as a target or tool in cancer immunotherapy research.

• Antimicrobial Activity: CXCL14 exhibits broad-spectrum antimicrobial effects against Gram-positive and Gram-negative bacteria, as well as fungi, by disrupting microbial membranes and binding bacterial DNA. Recombinant CXCL14 can be used to study innate immune responses and host-pathogen interactions.

• Muscle Biology: Recent studies show that CXCL14 promotes skeletal muscle mass growth and counteracts muscle atrophy induced by inflammatory or catabolic stimuli, making it relevant for research into sarcopenia, cachexia, and muscle wasting disorders.

• Neurodevelopment: CXCL14 is implicated in neuronal development and may influence neuroimmune interactions, providing a basis for studies in neurobiology and neuroinflammation.

• Metabolic Regulation: CXCL14 has been linked to body weight control and obesity phenotypes in mouse models, suggesting roles in metabolic research.

• Experimental Flexibility: Recombinant CXCL14 is validated for use in functional assays, ELISA, and cell-based studies, ensuring reproducibility and specificity in experimental design.

In summary, using recombinant Mouse CXCL14 enables targeted, mechanistic studies across immunology, oncology, microbiology, muscle physiology, neurobiology, and metabolism, supporting both basic research and translational applications.

Suitability for ELISA Standards

Recombinant mouse CXCL14 can be used as a standard for quantification and calibration in ELISA assays, but with important considerations regarding its specific application and formulation.

Recommended Use Cases

Recombinant CXCL14 proteins are well-suited for use as ELISA standards when properly formulated. The protein is typically expressed in E. coli and purified to high levels (>95% purity by SDS-PAGE), making it suitable for generating standard curves. Many commercial ELISA kits utilize recombinant mouse CXCL14 as their calibration standard, with typical standard curve ranges spanning from 0.16 to 10 ng/mL or higher depending on the assay platform.

Critical Formulation Considerations

The formulation of your recombinant protein is essential for ELISA applications. For use as an ELISA standard, recombinant CXCL14 should be obtained with bovine serum albumin (BSA) or other appropriate carrier proteins. This formulation choice is important because:

• BSA-containing formulations provide stability and prevent non-specific adsorption to plate surfaces
• Tag-free versions are available and maintain biological activity
• The carrier protein helps maintain protein integrity throughout the assay

Important Limitation

A critical restriction exists for certain recombinant preparations: ELISA standard recombinant proteins are not recommended for bioassay usage, as they are not tested for these applications. This means if your recombinant CXCL14 is specifically labeled as an "ELISA standard," it should be used only for quantification purposes in sandwich ELISA assays, not for functional bioassays involving cell stimulation or activation studies.

Best Practice Recommendation

For optimal results, verify that your recombinant mouse CXCL14 is formulated specifically for ELISA applications and contains appropriate stabilizing agents. This ensures reliable standard curve generation and accurate quantification of endogenous CXCL14 in your samples.

Recombinant Mouse CXCL14 has been validated for several key applications in published research, including functional assays, ELISA, and Western blot. In addition, it has been used in diverse biological studies to investigate its roles in immune modulation, muscle physiology, and metabolic regulation.

Validated Applications:

• Functional Assays: Recombinant Mouse CXCL14 is frequently used to assess its biological activity, such as chemotaxis of immune cells (e.g., monocytes, dendritic cells), modulation of T-cell responses, and effects on muscle cell growth and atrophy.
• ELISA (Enzyme-Linked Immunosorbent Assay): Used for quantifying CXCL14 protein levels in biological samples and validating its presence or activity.
• Western Blot: Applied to detect and confirm the expression of CXCL14 in cell lysates or tissue extracts.

Published Research Applications:

• Tumor Immunology: CXCL14 has been used to study its role in tumor suppression, particularly through recruitment and activation of CD8⁺ T cells and restoration of MHC-I antigen presentation in mouse models of HPV-associated cancers. It is also implicated in dendritic cell maturation and antitumor immunity.
• Muscle Biology: Recombinant CXCL14 has been shown to promote skeletal muscle mass growth and attenuate muscle atrophy in both in vitro (C2C12 myotubes) and in vivo (mouse tibialis anterior muscle) models. It reverses LPS- and dexamethasone-induced muscle wasting and modulates key signaling pathways related to muscle health.
• Metabolic Regulation and Obesity: Studies have used recombinant CXCL14 to investigate its involvement in body weight control, feeding behavior, and obesity phenotypes in mouse models, including leptin-deficient and Ay mutant mice.
• Chemotaxis and Immune Cell Recruitment: CXCL14 acts as a chemoattractant for monocytes and macrophage precursors, with activity demonstrated in dose-dependent migration assays.

Additional Notes:

• Recombinant Mouse CXCL14 is typically expressed in E. coli for research use.
• Its activity has been validated in both mouse and human cell systems, reflecting high sequence similarity and conserved function.

Summary Table:

These applications are supported by multiple peer-reviewed studies and are considered standard for recombinant CXCL14 research in mouse models.

To reconstitute and prepare Recombinant Mouse CXCL14 protein for cell culture experiments, dissolve the lyophilized protein in sterile distilled water or sterile PBS to a concentration of 0.1–1.0 mg/mL. For optimal stability and to prevent adsorption, it is recommended to include 0.1% BSA in the buffer.

Step-by-step protocol:

• Centrifuge the vial briefly (6000–10,000 rpm for 30 seconds) to collect all material at the bottom before opening.
• Add sterile distilled water or PBS to achieve the desired concentration (typically 0.1–1.0 mg/mL).
• If using for cell culture, include 0.1% BSA in the buffer to stabilize the protein and prevent loss due to adsorption to plastic surfaces.
• Gently mix by pipetting up and down or by gentle vortexing. Avoid vigorous agitation to prevent protein denaturation.
• Allow the solution to sit at room temperature for 15–30 minutes to ensure complete dissolution.
• Aliquot the stock solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at −20°C or −70°C for long-term storage, or at 2–8°C for short-term use (up to one month).

Additional notes:

• For some applications, adding 5–50% glycerol can further stabilize the protein during storage.
• Always use sterile technique to avoid contamination, especially for cell culture applications.
• Before use in cell culture, dilute the stock solution to the working concentration in your cell culture medium, ensuring the final BSA concentration is compatible with your assay.

Summary Table:

This protocol ensures protein stability and bioactivity for cell culture experiments.