Recombinant Mouse Cardiotrophin-1

Recombinant Mouse Cardiotrophin-1 (CT-1) is a multifunctional cytokine widely used in research for its roles in cardiac biology, neuroprotection, metabolism, and immunomodulation. Its recombinant form ensures high purity, reproducibility, and biological activity for experimental applications.

Key scientific reasons to use Recombinant Mouse Cardiotrophin-1 in research:

• Cardiac Differentiation and Remodeling: CT-1 promotes cardiomyocyte differentiation from mouse induced pluripotent stem cells via the JAK2/STAT3/Pim-1 signaling pathway, making it valuable for studies on cardiac development and regeneration. It induces reversible, physiologic hypertrophy and stimulates angiogenic signals, mimicking beneficial cardiac adaptation seen with exercise and improving cardiac function in disease models.

• Metabolic Regulation: CT-1 is a key regulator of energy metabolism, with demonstrated effects on fatty acid metabolism and insulin resistance. It has potential applications in obesity and metabolic syndrome research.

• Neuroprotection and Muscle Disease: CT-1 administration improves mitochondrial function and provides neuroprotection in mouse models. It has shown therapeutic benefits in motor neuron disease and spinal muscular atrophy models, improving functional and morphological outcomes.

• Immunomodulation: CT-1 acts as an anti-inflammatory cytokine and promotes IL-4-induced M2 macrophage polarization, relevant for studies on immune response and inflammation.

• Experimental Consistency: Recombinant CT-1 offers batch-to-batch consistency, defined activity (e.g., ED50 in cell proliferation assays), and suitability for in vitro and in vivo studies, ensuring reliable experimental outcomes.

Typical research applications include:

• Cardiac physiology and pathology models
• Stem cell differentiation protocols
• Metabolic disease studies
• Neurodegeneration and muscle disease models
• Immunology and inflammation assays

Summary:
Use recombinant mouse CT-1 to investigate cardiac differentiation, remodeling, metabolism, neuroprotection, and immunomodulation with high experimental reproducibility and defined biological activity.

Yes, recombinant Mouse Cardiotrophin-1 can generally be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose in your specific assay system. Most commercial ELISA kits for mouse Cardiotrophin-1 (CT-1) are designed to detect both natural and recombinant forms of the protein, and recombinant standards are commonly used for generating standard curves in quantitative ELISA protocols.

Key considerations and supporting details:

• Assay Compatibility: Many ELISA kits explicitly state that they recognize both natural and recombinant mouse CT-1, and their standard curves are often generated using recombinant protein. For example, one kit notes: "The assay will exclusively recognize both natural and recombinant Ms CT-1". Another development kit specifies it is suitable for measuring both forms.

• Standard Preparation: Guidelines for ELISA standard preparation recommend using a purified or recombinant protein to generate the standard curve, as this ensures accuracy and reproducibility. Recombinant proteins are widely accepted for this purpose, especially when native protein is not readily available.

• Validation: It is essential to confirm that the recombinant protein you intend to use is similar in sequence, folding, and post-translational modifications (if relevant) to the native protein detected by your assay antibodies. Most commercial recombinant mouse CT-1 proteins are produced in E. coli and are non-glycosylated, which is generally acceptable for standard curve generation, but you should verify compatibility with your specific ELISA kit.

• Kit-Specific Recommendations: Some ELISA kit manufacturers recommend using their supplied recombinant standard for best results, as it is validated for their antibody pairs and assay conditions. If you are using a custom or in-house ELISA, ensure your recombinant standard is well-characterized and that its concentration is accurately determined.

• Potential Limitations: Rarely, certain ELISA kits may not detect all recombinant forms due to differences in folding or epitope presentation, especially if the recombinant protein is truncated or modified. One source notes, "we can not guarantee the kit could detect recombinant protein" for all preparations, so empirical validation is advised.

Best Practices:

• Use a recombinant Mouse Cardiotrophin-1 standard that is as close as possible in sequence and structure to the native protein.
• Validate the standard curve by running serial dilutions of the recombinant protein in the same matrix as your samples (e.g., serum, plasma, buffer).
• Confirm linearity, recovery, and parallelism between the standard curve and endogenous samples to ensure accurate quantification.

In summary: Recombinant Mouse Cardiotrophin-1 is widely used and accepted as a standard for ELISA quantification, but always confirm compatibility with your specific assay and validate performance as part of your experimental setup.

Validated Research Applications

Recombinant mouse cardiotrophin-1 (CT-1) has been validated across multiple research applications in published studies, spanning cardiovascular biology, cellular differentiation, and metabolic research.

Cardiomyocyte Differentiation and Maturation

Recombinant CT-1 has been demonstrated to enhance cardiogenesis from mouse induced pluripotent stem cells (miPSCs) and facilitate the maturation of induced pluripotent stem cell-derived cardiomyocytes (iPS-CMs) through the JAK2/STAT3/Pim-1 signaling pathway. This application is particularly valuable for generating functional cardiac tissue for disease modeling and regenerative medicine approaches.

Cardiac Protection and Stress Response

The protein has been validated for studying cardioprotective mechanisms in cultured neonatal cardiac myocytes, where pretreatment with CT-1 protected cells against thermal stress (heat shock) and simulated ischemia/hypoxia conditions. These protective effects were associated with enhanced expression of heat shock proteins hsp70 and hsp90. Additionally, CT-1 has been shown to reduce apoptotic cell death induced by serum removal and ischemic stress, demonstrating its utility in investigating cell survival pathways.

Cardiac Remodeling and Hemodynamic Studies

Recombinant CT-1 has been used to study beneficial myogenic and vascular remodeling of the heart. In vivo studies have employed intravenous injection of CT-1 to examine dose-dependent effects on cardiac output, heart rate, mean arterial pressure, and systemic vascular resistance.

Metabolic Research

The protein has been validated in studies examining energy metabolism, where treatment of obese and diabetic mice with CT-1 increased energy expenditure and reduced food intake, correcting obesity and diabetes phenotypes.

Biomarker and Disease Association Studies

Recombinant CT-1 has been utilized in research investigating its role as a biomarker for left ventricular hypertrophy and dysfunction in hypertensive heart disease, as well as its association with cardiac fibrosis through the CT-1/galectin-3 axis in heart failure models.

To reconstitute and prepare Recombinant Mouse Cardiotrophin-1 (CT-1) protein for cell culture experiments, dissolve the lyophilized protein in sterile water or 10 mM PBS (pH 7.4) to a final concentration of 0.1–1.0 mg/mL. Do not vortex the solution; gently mix by pipetting or slow inversion.

Essential steps and best practices:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Use sterile technique throughout to avoid contamination.
• If the protein is sensitive or for extended storage, add a carrier protein such as 0.1% BSA or HSA to the buffer after initial reconstitution. This helps stabilize the protein and prevents adsorption to tube walls.
• After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Short-term storage: Store the reconstituted solution at 2–8 °C for up to one week.
• Long-term storage: Store aliquots at –20 °C to –80 °C for several months to a year.
• For cell culture, dilute the protein further in your culture medium or buffer as needed, ideally containing a carrier protein if not already present.

Additional notes:

• Avoid vigorous mixing (e.g., vortexing) as this may denature the protein.
• If recommended by the supplier, glycerol (5–50%) may be added for additional stabilization, especially for freeze-thaw protection.
• Always check the product datasheet for specific instructions regarding buffer composition, pH, and additives, as these may vary depending on the recombinant protein’s formulation.

Summary protocol:

1. Briefly centrifuge the vial.
2. Add sterile water or PBS (pH 7.4) to achieve 0.1–1.0 mg/mL.
3. Gently mix, do not vortex.
4. Add carrier protein if required.
5. Aliquot and store appropriately.
6. Dilute to working concentration in cell culture medium before use.

This protocol ensures maximum protein stability and activity for cell culture experiments.