Recombinant Mouse Gas6

Recombinant mouse Gas6 is a valuable research tool for investigating multiple biological pathways and cellular processes. Here are the key reasons to incorporate it into your research:

Biological Activity and Signaling

Recombinant mouse Gas6 functions as a ligand for the TAM receptor family (Axl, Dtk/Tyro3, and Mer), triggering Gas6/Axl signaling that regulates cell proliferation, survival, and migration. This makes it essential for studying receptor tyrosine kinase signaling in various cell types, including fibroblasts, endothelial cells, neural cells, and vascular smooth muscle cells.

Diverse Cellular Effects

The protein demonstrates dual regulatory capabilities depending on context. It can promote cell proliferation and prevent apoptosis independent of its mitogenic activity, while also inducing cell adhesion and chemotaxis in specific cell types. Additionally, Gas6 can inhibit inflammatory cytokine production from monocytes, macrophages, and microglia, while simultaneously functioning as a pro-inflammatory molecule through platelet activation.

Disease Model Applications

Recombinant mouse Gas6 has proven utility in studying multiple disease states:

• Infectious disease: Administration of Gas6 recombinant protein to elderly mice prevented pathogenic translocation of Klebsiella pneumoniae from the gastrointestinal tract and significantly prolonged survival by enhancing intestinal tight junction barriers
• Sepsis and organ damage: Gas6 ameliorates sepsis-induced organ damage and reduces reactive oxygen species formation
• Respiratory infections: Gas6/Axl signaling attenuates alveolar inflammation in ischemia-reperfusion-induced acute lung injury
• Cancer immunology: The protein is applicable to studying AXL-mediated drug resistance and tumor cell behavior

Experimental Validation

The protein is well-characterized for bioassay applications with established dose-response relationships (ED₅₀ values of 20-100 ng/mL for Axl downregulation in prostate carcinoma cells). It has been validated for binding activity assays and demonstrates consistent performance across multiple research applications.

Yes, recombinant Mouse Gas6 protein can be used as a standard for quantification or calibration in ELISA assays, provided it is properly formulated and validated for this purpose. Recombinant standards are commonly used in quantitative ELISA protocols to generate standard curves, enabling accurate measurement of Gas6 concentrations in biological samples.

Essential context and best practices:

• Formulation: For use as an ELISA standard, recombinant Gas6 is often supplied with a carrier protein such as BSA to enhance stability and prevent adsorption losses. Carrier-free formulations may be suitable for some applications, but stability and recovery should be validated.

• Standard Curve Preparation: The recombinant Gas6 standard should be serially diluted in the same buffer or matrix as your samples to minimize matrix effects and ensure parallelism between standard and sample curves. Always follow the ELISA kit or protocol instructions for standard preparation.

• Validation: ELISA kits designed to quantify mouse Gas6 typically use recombinant Gas6 as the calibration standard and report accurate quantification of both recombinant and native Gas6 in biological samples. It is important to confirm that the recombinant standard is recognized equivalently to native Gas6 by the antibodies used in your assay.

• Storage and Handling: Recombinant standards should be stored at -20°C or -80°C after reconstitution to maintain activity and avoid repeated freeze-thaw cycles. Follow manufacturer or protocol recommendations for storage and handling.

• Recovery and Precision: Published ELISA kits report high recovery rates and low coefficients of variation when using recombinant Gas6 as a standard, indicating reliable quantification.

Additional considerations:

• Species specificity: Ensure the recombinant Gas6 matches the species of your samples (mouse) and is compatible with the antibodies in your ELISA system.
• Documentation: Record lot numbers and concentrations for reproducibility and traceability.

Summary Table: Recombinant Mouse Gas6 as ELISA Standard

In conclusion, recombinant Mouse Gas6 is widely accepted and validated as a standard for ELISA quantification, provided best practices in preparation, validation, and handling are followed.

Validated Applications in Published Research

Recombinant mouse Gas6 has been validated for several key applications across diverse research areas:

Bioassay and Cell-Based Studies

The protein has been extensively validated for bioassay applications, demonstrating its functional activity in cell culture systems. Specifically, it has been used to assess mitogenic effects on multiple cell types including fibroblasts, endothelial cells, neural cells, vascular smooth muscle cells, and tumor-derived cell lines. The protein's ability to prevent apoptosis and induce cell adhesion and chemotaxis has been characterized through these bioassay approaches.

Inflammatory Disease Models

Recent research has validated recombinant Gas6 in acute lung injury models, where it was administered intraperitoneally to evaluate its effects on lung inflammation. In these studies, the protein successfully reduced proinflammatory cytokine production, neutrophil infiltration, and inflammatory markers in alveolar macrophages. The Gas6/AXL signaling pathway has also been validated for studying sepsis-induced organ damage and tight junction injury in both in vitro and in vivo settings.

Binding Activity Assays

The protein has been validated for binding activity applications, confirming its capacity to interact with TAM family receptors (Axl, Dtk/Tyro3, and Mer). This has been demonstrated through its ability to downregulate Axl expression in cell culture systems.

In Vivo Applications

Recombinant Gas6 has been validated for in vivo studies examining sepsis-induced organ damage and reactive oxygen species formation, as well as investigations into microglia efferocytosis and anti-inflammatory responses in central nervous system disease models.

These applications collectively demonstrate the protein's utility in studying tissue homeostasis, immune regulation, and stress-induced cellular responses across multiple biological systems.

Reconstitution Procedures

Recombinant mouse Gas6 proteins are typically supplied in lyophilized form and require reconstitution before use in cell culture experiments. The specific reconstitution protocol depends on the formulation variant you are using.

Standard Reconstitution Protocol

For most recombinant mouse Gas6 preparations, reconstitute the lyophilized protein at a concentration of 100 μg/mL in sterile PBS. Some full-length variants may require reconstitution at 250 μg/mL in sterile PBS. Before opening the vial, briefly centrifuge the tube at 4°C to recover all contents and ensure complete recovery of the protein.

Alternative reconstitution approaches include using sterile distilled water at concentrations ranging from 0.1–0.5 mg/mL, depending on your specific application requirements. Gently shake or mix the vial to solubilize the protein completely rather than vigorous agitation.

Formulation Considerations

Carrier Protein Variants

Some formulations include BSA as a carrier protein, while carrier-free versions are also available. Carrier-free preparations may be preferred for certain downstream applications such as binding assays or receptor interaction studies. Carrier-free formulations are typically supplied in PBS with glycerol (25% glycerol, pH 7.2).

Purity and Quality Specifications

Recombinant mouse Gas6 proteins are typically supplied with purity greater than 95% as determined by SDS-PAGE. Endotoxin levels are generally less than 0.1–1 EU/μg, making these preparations suitable for cell culture work.

Storage and Stability

Optimal Storage Conditions

Store reconstituted or lyophilized protein at temperatures below -20°C in a manual defrost freezer. The protein remains stable for approximately 6 months after receipt when stored properly.

Critical Handling Practices

Minimize freeze-thaw cycles, as repeated freezing and thawing can compromise protein integrity and biological activity. If you require frequent access to the protein, consider preparing aliquots in appropriate volumes to reduce the number of freeze-thaw events.

Preparation for Cell Culture Applications

Once reconstituted, the Gas6 protein is ready for use in bioassays and cell culture experiments. Gas6 functions as a mitogen for fibroblasts, endothelial cells, neural cells, and vascular smooth muscle cells, and also prevents apoptosis independent of its mitogenic activity. The protein can induce cell adhesion and chemotaxis in specific cell types.

For experiments involving TAM receptor signaling (Axl, Dtk/Tyro3, and Mer), ensure that the reconstituted Gas6 maintains proper post-translational modifications, particularly γ-carboxylation of the Gla domain, which is essential for receptor binding. If using recombinant protein produced in mammalian expression systems, this modification is typically already present in commercial preparations.