Antibody System Human sCD14 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 sCD14 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 sCD14 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 sCD14 is introduced into the wells, anchoring onto an alternative epitope of the captured sCD14 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 sCD14 bound in the initial step. The reaction is terminated with a stop solution, and absorbance is quantified spectrophotometrically.

Key Features and Performance Metrics

• Refined Sensitivity Profile: Exhibits an analytical sensitivity down to 27.26 pg/mL, facilitating the accurate tracking of low-level sCD14 concentrations in diverse biological samples.

• Linear Dynamic Range: Delivers a reliable quantitative range tracking from 93.75 pg/mL to 6,000 pg/mL to accommodate broad sample variation.

• 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 = 2745.4 pg/mL, SD = 150.1, CV = 5.5%

    • Sample 2: Mean = 642.4 pg/mL, SD = 66.8, CV = 10.4%

    • Sample 3: Mean = 159.6 pg/mL, SD = 7.0, CV = 4.4%

  • 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 = 2800.2 pg/mL, SD = 279.6, CV = 10.0%

    • Sample 2: Mean = 747.4 pg/mL, SD = 66.9, CV = 9.0%

    • Sample 3: Mean = 161.7 pg/mL, SD = 7.4, CV = 4.6%

Technical Specifications

• Catalog No.: KHC33701

• Assay Configuration: Quantitative Sandwich Enzyme Immunoassay

• Targeted Biomarker: Human Soluble CD14 (sCD14)

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

• Detection Method: Colorimetric

• Assay Range: 93.75 to 6,000 pg/mL

• Analytical Sensitivity: 27.26 pg/mL

• Solid-Phase Target: Anti-Human sCD14 Specific Antibody

• Detection Secondary Layer: Biotin-Anti-Human sCD14 / Streptavidin-HRP Conjugate

Alternative Names: Monocyte differentiation antigen CD14, Myeloid cell-specific leucine-rich glycoprotein, CD14.

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 6,000 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 over a standard six-month framework, demonstrating an activity loss rate of less than 20 percent when kept at the manufacturer-specified storage parameters.

• 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.