5(6)-Aminofluorescein (5(6)-AFM) is a high-purity, high-quality chemical utilized as a molecular probe and a significant fluorescent agent.

Instructions
1. Synthesis of 5(6)-FITC:
a. Reaction process: 5(6)-AFM is used as an amine precursor to react with isothiocyanate to generate 5(6)-FITC. The reaction is generally carried out in an alkaline environment. Isothiocyanate (FITC) is added to the reaction to react with 5(6)-AFM to form a stable fluorescent dye.
b. Reaction conditions: The reaction is usually carried out in an aqueous phase or an organic solvent (such as DMF). The reaction temperature is generally room temperature or slightly higher. The reaction time is 1-2 hours, and then the unreacted substances are removed by precipitation, filtration or washing.
2. Labeling proteins or antibodies:
The generated 5(6)-FITC can be used to label molecules with amino groups (such as proteins, antibodies, peptides, etc.). The labeling process usually includes:
a. Dissolve 5(6)-FITC in a suitable solvent (such as DMSO, PBS buffer, etc.).
b. React with the target molecule (such as protein), usually under appropriate pH conditions.
c. After the reaction is complete, unreacted dye is removed by dialysis or column chromatography.
3. Fluorescence detection:
Molecules labeled with 5(6)-FITC can be imaged and quantitatively analyzed by fluorescence microscopy or flow cytometry.
The excitation wavelength of 5(6)-FITC is 490 nm and the emission wavelength is 520 nm, which is suitable for most fluorescence imaging and analysis platforms.