Universal Nucleic Acid ELISA Kit

The Universal Nucleic Acid ELISA Kit provides a plate-based method for colorimetric detection and calibration-based quantification analysis of dual-labeled nucleic acids generated from amplification reactions. Hapten-labeled nucleic acids (FAM/FITC or Digoxigenin) are captured on antibody-coated microplates and detected using a biotin–streptavidin HRP system with TMB substrate. Following stop-solution addition, absorbance is measured using a standard microplate reader to quantify relative nucleic acid levels.

This kit offers a practical endpoint alternative to gel electrophoresis or probe-based fluorescence detection for amplification workflows.

Research Use Only. Not for diagnostic procedures.

Assay Principle

The assay functions as a dual-labeled sandwich ELISA:

• Hapten-labeled nucleic acids (FAM/FITC or Digoxigenin) generated via modified primers bind to immobilized anti-hapten antibodies.

• A biotin modification incorporated during amplification enables enzymatic detection using streptavidin-HRP.

• HRP-mediated TMB conversion produces a colorimetric signal proportional to nucleic acid concentration, read at 450 nm after stop-solution addition.

Applications

• Endpoint detection of PCR, RT-PCR, LAMP, and RPA products

• Amplification optimization and comparative analysis

• DNA-binding or hybridization assays

• Enzymatic activity and inhibition screening

• Nuclease activity testing and assay development

Performance Characteristics

• Capture Formats: Anti-FAM/FITC (Kit 01) or Anti-Digoxigenin (Kit 02)

• Dynamic Detection Range:

  • FAM/FITC: 10–760 attomoles/µL

  • Digoxigenin: 10–920 attomoles/µL
    (50 µL sample volume)

• Detection: Streptavidin-HRP with TMB substrate

• Readout: Absorbance at 450 nm

• Format: Standard 96-well ELISA workflow

• Plates optimized for high-efficiency nucleic acid capture and signal performance

Kit Contents

• Antibody-coated 96-well microplate

• Assay buffers and wash solutions

• Streptavidin-HRP conjugate

• TMB substrate and stop solution

• Protocol documentation

Required Materials (Not Included)

• Dual-modified primers (hapten + biotin)

• Amplified nucleic acid samples

• Microplate reader capable of 450 nm detection

• Standard ELISA washing supplies

Features & Benefits

• Converts amplification products into a stable ELISA readout

• No fluorescent probes or electrophoresis required

• Compatible with common amplification platforms

• Uses standard ELISA instrumentation

• Suitable for high-throughput assay development and screening

• Cost-effective endpoint analysis workflow