0
Invitrogen™ FluoReporter™ Biotin Quantitation Assay Kit, for biotinylated proteins
Catalog No :
CAS Number :
Brand :
In Stock
Specifications:
| Application | Fluorescence-Based Assay | ||
| Storage Temperature | -5 to -30°C | ||
| Product Type | Molecular Biology Reagent | Forms | Kit with Various components |
| Product Brand | Thermo Fisher Scientific™, Invitrogen | ||
| Product Grade | Molecular Biology | ||
The Invitrogen™ FluoReporter™ Biotin Quantitation Assay Kit is a highly sensitive, fluorescence-based solution designed to determine the degree of biotinylation in nucleic acids and proteins. This assay uses Biotective™ Green reagent, which fluoresces upon displacement of quencher ligands by biotin, allowing precise, quantitative detection. With up to 50-fold greater sensitivity than HABA dye-based methods, the kit enables reliable analysis using standard microplate readers compatible with FITC fluorescence.
Key Features and Benefits
Exceptional Sensitivity
- Detects 4–80 picomoles of biotin per sample, offering excellent performance even in low-biotinylation contexts.
- Requires as little as 13 ng of labeled nucleic acid, depending on the extent of labeling.
Fluorescence-Based Detection
- Biotective™ Green reagent emits fluorescence upon biotin displacement.
- Excitation/Emission Maxima: 495/519 nm – compatible with standard FITC filter sets.
Broad Compatibility
- Applicable to biotinylated proteins and nucleic acids, including samples labeled with NHS-biotin or sulfo-NHS-biotin.
- Positive control and standard included to validate assay performance and enable standard curve generation.
Streamlined, Reliable Workflow
- Designed for easy integration into research workflows requiring labeling efficiency verification or assay validation.
- Use with standard fluorescence microplate readers in a 96-well format.
Applications
- Quantifying biotin on cDNA, RNA, or DNA in molecular biology workflows
- Measuring protein biotinylation levels for ELISA, Western blot, or immunoprecipitation
- Assessing labeling efficiency in antibody conjugation
- Optimizing biotin-streptavidin or biotin-avidin-based affinity assays
Technical Specifications
| Property | Details |
|---|---|
| Catalog Number | F30751 |
| Detection Method | Fluorescence (Biotective™ Green) |
| Excitation/Emission | 495/519 nm (FITC-compatible) |
| Detection Range | 4–80 pmol biotin per sample |
| Minimum Sample Amount | ≥13 ng biotinylated nucleic acid (dependent on labeling) |
| Label Target | Proteins, Nucleic Acids |
| Label Type | Biotin |
| Product Format | 1 Kit |
| Storage Conditions | –5 to –30°C; protect from light |
| Shipping Condition | Room temperature |
| Instrument Compatibility | Fluorescence microplate reader (96-well, FITC channel) |
Kit Contents
| Component | Description |
|---|---|
| Component A | Biotective™ Green reagent (lyophilized), 5 vials |
| Component B | Biocytin (200 μM), 100 μL |
| Component C | Protease (lyophilized), 5 × 5 U (Note A) |
| Component D | 10X Phosphate-buffered saline (PBS), 4 mL |
| Component E | Biotinylated goat anti–mouse IgG, 2 mg/mL, 37.5 μL (Note B) |
Note A: Protease is required to release biotin from labeled macromolecules for detection.
Note B: Biotinylated antibody serves as a positive control and protein standard.
Storage and Handling
- Store the kit at –5 to –30°C and protect all reagents from light.
- This kit supports 5 independent assays, each using 8 wells in triplicate for standard curves and 3 dilutions of the sample in triplicate, totaling 33 wells per assay.
- To conserve materials, fewer wells may be used per run. A single standard curve may be used across multiple samples in the same experimental session.
- Biocytin (a biotinylated lysine analog) is used as a standard as it mimics the form of biotin released after proteolysis.
- The included biotinylated goat anti–mouse IgG provides a reliable positive control.
The FluoReporter™ Biotin Quantitation Assay Kit is a sensitive, easy-to-use solution for quantifying biotinylation in nucleic acids and proteins. With ready-to-use reagents, broad compatibility, and robust fluorescence-based detection, this kit is ideal for researchers needing reliable biotin quantification in molecular biology, immunology, and bioconjugation workflows.