Fluorescein Isothiocyanate (FITC), 5- and 6-Isomer Mixture provides an amine-reactive fluorescent dye for bioconjugation and molecular labeling research. This reagent contains both positional isomers of fluorescein isothiocyanate, while its isothiocyanate functionality supports covalent labeling of proteins, antibodies, peptides, and other primary amine-containing molecules.
Key Features and Benefits
- 5- and 6-Isomer Mixture: Contains two positional isomers of fluorescein isothiocyanate for fluorescence labeling and bioconjugation applications.
- Reactive Isothiocyanate Group: Reacts with compatible primary amines to form stable thiourea linkages during covalent labeling.
- Green Fluorescence: Provides bright green fluorescence with typical excitation near 495 nm and emission near 520 nm.
- Protein Labeling Capability: Enables fluorescent modification of proteins through accessible lysine residues and other primary amine groups.
- Antibody Conjugation Compatibility: Supports preparation of fluorescent antibody conjugates for immunoassays, microscopy, and molecular detection research.
- Established Fluorescein Chemistry: Uses a widely recognized fluorescent scaffold compatible with many standard fluorescence detection systems.
- Covalent Labeling Stability: Forms thiourea bonds that retain the fluorescent label on compatible biomolecules under suitable conditions.
- Bioconjugation Versatility: Enables labeling of peptides, proteins, antibodies, and other molecules containing accessible primary amines.
- Analytical Detection Potential: Supports fluorescence-based detection, molecular tracking, and quantitative assay development.
Applications
- Protein Fluorescent Labeling: Enables covalent attachment of fluorescein labels to proteins through reactions with accessible primary amine groups.
- Antibody Conjugation: Supports preparation of FITC-labeled antibodies for immunofluorescence, immunoassays, and molecular detection studies.
- Flow Cytometry: Enables development of fluorescent antibody conjugates and cellular probes for compatible flow cytometry applications.
- Immunofluorescence Microscopy: Supports fluorescent labeling of antibodies and biomolecules for investigating cellular structures and protein localization.
- Cellular Imaging Research: Enables preparation of fluorescent probes for studying cellular binding, uptake, and molecular interactions.
- Peptide Labeling: Supports conjugation with compatible amine-containing peptides for fluorescence tracking and biochemical studies.
- Fluorescence-Based Assays: Enables development of analytical assays for detecting labeled biomolecules and evaluating binding interactions.
- Biomolecular Interaction Studies: Supports preparation of fluorescent conjugates for investigating protein binding, molecular recognition, and biochemical interactions.
- Fluorescent Probe Development: Provides an amine-reactive dye for creating experimental fluorescent probes and chemically modified biomolecules.
Storage and Handling
- Storage: Store at −20°C in a dry, tightly sealed container, protected from light and moisture. Follow product-specific storage instructions when available. Minimize repeated temperature changes and prolonged exposure to humidity.
- Handling: Handle using clean, dry equipment and protect the dye from excessive light exposure. Prepare concentrated stock solutions in anhydrous DMSO or DMF when appropriate. Confirm solubility and solvent compatibility before beginning the labeling procedure.
- React FITC with accessible primary amines on compatible proteins, antibodies, peptides, or other biomolecules. Use an appropriate amine-free buffer, such as bicarbonate or carbonate buffer, under mildly alkaline conditions. Avoid Tris and other primary amine-containing buffers because they compete with the target molecule.
- Optimize the dye-to-biomolecule molar ratio, reaction time, and buffer conditions for the intended application. Isothiocyanate groups react with primary amines to form thiourea linkages. Protect reaction mixtures from unnecessary light exposure and avoid prolonged incubation under harsh conditions.
- Remove unreacted dye through dialysis, size-exclusion chromatography, or another suitable purification method. Evaluate labeling efficiency, degree of labeling, fluorescence intensity, and conjugate stability using appropriate analytical techniques. Excessive labeling may affect biomolecule activity, solubility, or fluorescence behavior. Confirm the product’s spectral properties, purity, and isomer composition before use.
Fluorescein Isothiocyanate (FITC), 5- and 6-Isomer Mixture combines green fluorescence with amine-reactive isothiocyanate chemistry for covalent biomolecule labeling and specifically supports antibody conjugation, protein labeling, flow cytometry, fluorescence microscopy, and biochemical assay development.
AxisPharm offers 5000+ PEG Linkers with high purity. Different kinds of PEG Reagents may be available by custom synthesis.
