Thermo Scientific™ Pierce™ Fluorescence Biotin Quantitation Kit

The Thermo Scientific™ Pierce™ Fluorescence Biotin Quantitation Kit is a sensitive and efficient microplate-based assay designed to accurately determine the biotinylation level of antibodies and other biomolecules using just 10 μL of sample. Utilizing a DyLight-labeled avidin-HABA complex, this assay delivers high sensitivity and reproducibility, making it suitable for labs engaged in antibody labeling, protein conjugation, or any workflow requiring precise control of biotin incorporation.

Key Features & Benefits

• High Sensitivity
  Detects 10 to 60 picomoles of biotin, enabling accurate measurements even with low-concentration samples.
• Minimal Sample Requirement
  Requires only 10 µL of sample, a fraction of what is needed for colorimetric or competing fluorescent assays.
• Fast Turnaround
  The entire assay is completed in just 5 minutes, minimizing hands-on time and increasing throughput.
• Superior Accuracy
  Generates reliable, linear standard curves using the provided biocytin control, ensuring precise biotin-to-protein ratio calculations.
• Optimized for Fluorescence Detection
  Utilizes a fluorescent avidin-HABA reporter that increases signal upon biotin displacement. Readout is measured at excitation 494 nm / emission 520 nm, compatible with standard fluorescence microplate readers.
• Improved Over Colorimetric Methods
  Unlike the traditional HABA colorimetric assay, this fluorescent version is significantly more sensitive and uses 90% less sample volume.

How It Works

The assay is based on the strong, specific interaction between avidin and biotin. In this fluorescence method, HABA (4'-hydroxyazobenzene-2-carboxylic acid), a weakly-binding dye, is pre-complexed with avidin (DyLight Reporter). When biotin in the sample is introduced, it displaces the HABA dye, resulting in fluorescence enhancement that is directly proportional to the amount of biotin present.

Kit Contents

• Sufficient for: Up to 216 fluorescence-based samples
• Required but not included:
  • Black opaque 96-well microplate
  • Fluorescence plate reader (Ex 494 nm / Em 520 nm)

Applications

• Measuring biotinylation efficiency of antibodies, enzymes, or other proteins
• Determining biotin-to-protein ratios post-labeling
• Standardizing biotin-conjugate concentrations for downstream streptavidin-based assays
• Quality control in conjugation and assay development workflows

Storage Conditions

• Store at 4°C
• Do not freeze
• Shipped at ambient temperature

The Pierce Fluorescence Biotin Quantitation Kit offers a robust, rapid, and highly sensitive method for determining the degree of biotinylation in biomolecules. Its low sample requirement, superior accuracy, and compatibility with standard fluorescence equipment make it an essential tool for researchers working with biotinylated antibodies and proteins in diagnostics, proteomics, and assay development.