Recombinant Cardiac Troponin I/T/C Complex

Using Recombinant Cardiac Troponin I/T/C Complex (24103P) in research applications is recommended for its high purity, defined composition, and relevance as a standard or calibrator in cardiac biomarker assays, particularly for studies involving myocardial injury and cardiac muscle physiology.

Key scientific advantages and applications include:

• Biomarker Standardization: The troponin complex (I/T/C) is the gold-standard biomarker for detecting cardiac injury, including acute myocardial infarction (AMI), due to its cardiac-specific isoforms and high diagnostic sensitivity. Recombinant complexes provide a consistent, well-characterized standard for calibrating immunoassays and validating diagnostic platforms.

• Defined Molecular Composition: Recombinant production ensures the complex contains the correct stoichiometry of cardiac troponin I, T, and C subunits, mimicking the native cardiac troponin complex. This is critical for assay specificity and for studying protein-protein interactions relevant to cardiac muscle contraction.

• High Purity and Quality Control: The recombinant complex is typically purified to >90–95% purity (as determined by RP-HPLC and SDS-PAGE), minimizing background interference and ensuring reproducibility in quantitative assays and biochemical studies.

• Versatility in Research Applications:

  • Immunoassay Calibration: Used as a calibrator or control in ELISA, Western blot, and other immunoassays for cardiac troponin detection.
  • Functional Studies: Enables investigation of the molecular mechanisms of cardiac muscle contraction, calcium sensitivity, and the effects of post-translational modifications on troponin function.
  • Drug Screening and Toxicology: Facilitates studies on drug-induced cardiotoxicity and myocardial injury by providing a reliable standard for troponin release assays.
  • Cellular and Biochemical Assays: Suitable for cell culture, activity assays, and protein-protein interaction studies due to its recombinant origin and defined structure.

• Species and Isoform Specificity: Recombinant complexes are typically derived from human cardiac isoforms, ensuring relevance for translational research and minimizing cross-reactivity with skeletal muscle troponins.

• Batch-to-Batch Consistency: Recombinant production in E. coli or other expression systems allows for scalable manufacturing and consistent quality across lots, which is essential for reproducible research outcomes.

In summary, Recombinant Cardiac Troponin I/T/C Complex (24103P) is a scientifically robust choice for research requiring precise, reproducible, and cardiac-specific troponin standards, especially in the context of myocardial injury diagnostics, cardiac physiology, and assay development.

Yes, you can use recombinant Cardiac Troponin I/T/C Complex (24103P) as a standard for quantification or calibration in ELISA assays, provided it is appropriately validated for your specific assay system. This type of recombinant troponin complex is commonly used as a calibrator or standard in research and diagnostic immunoassays targeting cardiac troponins.

Key considerations and supporting details:

• Intended Use: Recombinant cardiac troponin ITC complexes are specifically produced and characterized for use as standards and calibrators in immunoassays, including ELISA. The product you referenced (24103P) is described as having >90% purity and is formulated for laboratory use.

• Validation: While recombinant complexes are suitable as standards, you must ensure that the recombinant form is recognized equivalently by the antibodies used in your ELISA. Differences in post-translational modifications, folding, or epitope presentation between recombinant and native proteins can affect assay performance. It is recommended to run a standard curve and compare recovery and linearity with known controls.

• Commutability: For clinical or diagnostic applications, the standard should be commutable—that is, it should behave similarly to native troponin in patient samples across different assay platforms. For research use, this is less critical, but for quantitative accuracy, it is still important to verify.

• Concentration Determination: The concentration of the recombinant complex is typically determined by spectrophotometry (A280) using a defined extinction coefficient. Ensure accurate dilution and handling to maintain calibration integrity.

• Formulation: Some recombinant troponin complexes are supplied in buffers containing reducing agents (e.g., β-mercaptoethanol), which may affect certain assay formats. Confirm compatibility with your ELISA reagents and protocol.

• Precedent: Recombinant troponin subunits and complexes are widely used as standards in commercial ELISA kits and have been validated in peer-reviewed studies for this purpose.

Best Practices:

• Prepare a standard curve using serial dilutions of the recombinant complex in the same matrix as your samples (e.g., serum, plasma, or assay buffer).
• Validate the standard curve for linearity, sensitivity, and reproducibility within your assay system.
• If possible, compare results with a reference material or native troponin standard to assess equivalence.

Summary Table: Use of Recombinant Cardiac Troponin Complex as ELISA Standard

In summary: Recombinant Cardiac Troponin I/T/C Complex (24103P) is suitable as a standard for ELISA quantification, but you must validate its performance in your specific assay context to ensure accurate and reliable results.

The Recombinant Cardiac Troponin I/T/C Complex (24103P) has been validated for use in a variety of published research applications, primarily as a molecular tool for biochemical and immunological studies. Based on the available literature and product information:

• Immunoassay Standard/Calibrator: The recombinant complex is frequently used as a calibrator or standard in immunoassays for the detection and quantification of cardiac troponin I (cTnI) and cardiac troponin T (cTnT). This is supported by studies evaluating novel cardiac troponin assays, where recombinant troponin complexes are used for value assignment and assay calibration (Hytest, Leinco, QED Bioscience).

• Biochemical and Structural Studies: The complex is used in research investigating the structure, assembly, and function of the cardiac troponin complex. For example, recombinant troponin subunits and complexes are incorporated into reconstituted cTn complexes or cardiac thin filaments to study structure-function relationships and calcium sensitivity (Nature Communications, PMC articles).

• Immunological Applications: The recombinant complex is employed in the development and validation of immunochemical assays targeting different forms of the troponin complex, including intact and fragmented forms, which are relevant in monitoring myocardial injury (Hytest, Nature Communications).

• Western Blotting (WB): The product is validated for use in Western blotting applications, as indicated by QED Bioscience and other suppliers.

• Native Enrichment and Mass Spectrometry: Native nanoproteomics strategies have used recombinant or endogenous troponin complexes for enrichment and structural elucidation via native top-down mass spectrometry, directly from human heart tissue (Nature Communications).

In summary, the Recombinant Cardiac Troponin I/T/C Complex (24103P) has been validated for use as a calibrator in immunoassays, in biochemical and structural studies, immunological assays, Western blotting, and advanced proteomic analyses such as native mass spectrometry.

To reconstitute and prepare the Recombinant Cardiac Troponin I/T/C Complex (24103P) for cell culture experiments, follow these steps based on the most relevant technical data and best practices for recombinant troponin complexes:

1. Check the Protein Formulation and Storage Conditions

• This complex is typically supplied in a buffer containing 500 mM NaCl, 20 mM Tris-HCl, and 60 mM β-mercaptoethanol, pH 7.5.
• If the protein is provided as a liquid, it is ready for dilution and use.
• If provided lyophilized, reconstitution is required.

2. Reconstitution (if lyophilized)

• Centrifuge the vial briefly to collect all material at the bottom.
• Use the recommended buffer for reconstitution. For troponin complexes, a buffer such as 20 mM Tris-HCl, pH 7.5, with 500 mM NaCl and 60 mM β-mercaptoethanol is appropriate. If the datasheet suggests a different buffer (e.g., with urea for solubilization), follow that protocol.
• Add sterile buffer to achieve a final concentration of 0.1–1 mg/mL (typical for protein stock solutions).
• Gently mix by pipetting or slow inversion; avoid vigorous vortexing to prevent denaturation.

3. Aliquoting and Storage

• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Short-term storage (up to 2–4 weeks): 4°C.
• Long-term storage: -20°C or lower, in aliquots.
• Avoid more than 5 freeze-thaw cycles; activity loss is minimal up to this point.

4. Preparation for Cell Culture

• Before adding to cell culture, dilute the protein into your cell culture medium to the desired working concentration.
• If the storage buffer contains high salt or β-mercaptoethanol, consider buffer exchange or dialysis into a cell-compatible buffer (e.g., PBS or serum-free medium) to avoid cytotoxicity, as high β-mercaptoethanol and salt can be harmful to cells.
• Sterile filter the final solution (0.22 μm) if sterility is required for cell culture.

5. General Best Practices

• Always consult the lot-specific datasheet for any special instructions or deviations from standard protocols.
• Handle all recombinant proteins using standard laboratory safety procedures.

Summary Table: Key Steps

Note: If your application is sensitive to reducing agents or high salt, perform a buffer exchange before use in cell culture. Always verify compatibility with your specific cell line and experimental design.

These recommendations are based on standard protocols for recombinant troponin complexes and the specific formulation details for 24103P.