Antibody System Human IL6 ELISA Kit

Principle of the Assay

This assay utilizes a high-performance, solid-phase quantitative sandwich enzyme immunoassay technique to measure target protein levels:

• Target Capture: A monoclonal antibody specific for Human IL-6 has been uniformly pre-coated onto the solid-phase surface of a 96-well microplate.

• Specific Binding: Calibrated standards or unknown biological samples are pipetted directly into the microwells. Any endogenous Human IL-6 present in the sample matrix is securely bound and immobilized by the solid-phase capture antibody.

• Sandwich Conjugation: Unbound matrix elements are removed via a thorough wash cycle. A biotin-labeled secondary antibody specific for Human IL-6 is introduced into the wells, anchoring onto an alternative epitope of the captured IL-6 target to establish a stable sandwich complex.

• Enzyme Complexing: Following another wash step to clear away unbound detection antibody, Streptavidin conjugated to Horseradish Peroxidase (Streptavidin-HRP) is introduced to bind specifically to the biotinylated domains.

• Signal Amplification and Readout: After a final wash to remove excess enzyme reagent, a colorimetric substrate solution is added to initiate enzymatic signal development. Because this is a traditional sandwich format, the resulting color intensity develops in direct proportion to the concentration of Human IL-6 bound in the initial step. The reaction is terminated with a stop solution, and absorbance is quantified spectrophotometrically.

Key Features and Performance Metrics

• Ultra-Sensitive Detection Capability: Exhibits an elite analytical sensitivity down to 0.92 pg/mL, facilitating the accurate tracking of trace-level IL-6 concentrations.

• Refined Quantitative Range: Delivers a precise linear tracking range from 4.69 pg/mL to 300 pg/mL, optimized for high-resolution cytokine profiling.

• Rigorous Recovery Metrics: Validated with a target recovery range between 80 percent and 120 percent, indicating minimal matrix interference across verified human sample matrices.

• Exceptional Precision Profile:

  • Intra-Assay Precision (Precision within an assay): <10% Coefficient of Variation (CV). Validated across three reference samples evaluated sixteen times on a single plate:

    • Sample 1: Mean = 158.0 pg/mL, SD = 13.3, CV = 8.4%

    • Sample 2: Mean = 36.3 pg/mL, SD = 3.4, CV = 9.4%

    • Sample 3: Mean = 9.0 pg/mL, SD = 0.5, CV = 5.9%

  • Inter-Assay Precision (Precision between assays): <15% Coefficient of Variation (CV). Verified across three independent samples tested in twenty-four separate runs:

    • Sample 1: Mean = 153.5 pg/mL, SD = 16.6, CV = 10.8%

    • Sample 2: Mean = 34.1 pg/mL, SD = 3.3, CV = 9.7%

    • Sample 3: Mean = 8.9 pg/mL, SD = 1.0, CV = 10.7%

Technical Specifications

• Catalog No.: KHC15801

• Assay Configuration: Quantitative Sandwich Enzyme Immunoassay

• Targeted Biomarker: Human Interleukin-6 (IL-6)

• Validated Matrices: Serum, Plasma, and Cell Culture Supernatant

• Detection Method: Colorimetric

• Assay Range: 4.69 to 300 pg/mL

• Analytical Sensitivity: 0.92 pg/mL

• Solid-Phase Target: Anti-Human IL-6 Specific Antibody

• Detection Secondary Layer: Biotin-Anti-Human IL-6 / Streptavidin-HRP Conjugate

Applications and Workflow Summary

This sandwich ELISA platform is highly optimized for high-throughput cytokine tracking, acute-phase response monitoring, and systemic inflammation profiling. Investigators should observe the following guidelines:

• Dilution and Layout: Unknown serum, plasma, or supernatant matrices must be screened alongside the provided reference standards. Samples exhibiting target concentrations exceeding the maximum range of 300 pg/mL must be diluted empirically in standard assay matrix buffer prior to running the assay.

• Analysis Requirements: To achieve absolute quantification accuracy, investigators must generate a standard curve simultaneously during every plate run, plotting standard absorbance profiles against log-transformed concentration values.

Handling, Stability and Storage

• Kit Shipping: Distributed under secure, temperature-controlled conditions between 2 and 8 degrees Celsius to safeguard protein matrix and enzyme binding configurations.

• Operational Shelf Life: Built with high-grade components to guarantee consistent, low-decay signal retention, demonstrating an activity loss rate of less than 10 percent prior to the expiration date when kept under appropriate storage conditions.

• Storage Guidance: Keep the complete kit stored strictly at the recommended refrigeration temperature upon receipt. Ensure that unused microplate wells remain sealed within their original desiccant pouch to guard against moisture compromise.