Description
Fluorescein isothiocyanate (FITC) is an amine-reactive derivative of fluorescein dye that has wide-ranging applications as a label for antibodies and other probes, for use in fluorescence microscopy, flow cytometry and immunofluorescence-based assays such as Western blotting and ELISA. The isothiocyanate variety of fluorescein efficiently labels antibodies and other purified proteins at primary amines (lysine side chains)
Excitation source: 488 nm spectral line, argon-ion laser
Excitation wavelength: 494 nm
Emission wavelength: 518 nm
Extinction coefficient: > 70,000 M-1cm-1
Solubility: Soluble in aqueous buffers at pH > 6
Reactive groups: Isothiocyanate, reacts with primary amines at pH 7.0 to 9.0
Chemical Properties
Dark yellow powder, soluble in water and decomposed in water. It is easily soluble in alkaline aqueous solution, and the strong green fluorescence disappears and precipitates. After adding alkali, it dissolves and shows fluorescence. Insoluble in acetone, ether and petroleum ether. Hygroscopic.
Uses
Fluorescein Isothiocyanate is a fluorescent reagent with incompletely defined position of the NCS. Fluorescein Isothiocyanate is used in multifunctional uniform nanoparticles composed of a magnetite nanocrystal core and a mesoporous silica shell for magnetic resonance, fluorescence imaging. and drug delivery. Dyes and metabolites.
Uses
For fluorescent labeling of proteins (e.g. immunofluorescence);
H Protein labeling for gel chromatography.
Label antibodies for use as immunofluorescent probes.
Label oligonucleotides for hybridization probes.
Detect proteins in gels and on Western blots.
Application
Fluorescein isothiocyanate (FITC) is yellow-orange in color with an absorption maximum at 495nm. Upon excitation, it emits a yellow-green color with an emission maximum at 525nm.
It is widely used to attach a fluorescent label to proteins via the amine group. The isothiocyanate group reacts with amino terminal and primary amines in proteins. It has been used for the labeling of proteins including antibodies and lectins.
Fluorescein isothiocyanate isomer I has been proposed as a contact sensitizer.
Definition
ChEBI: Fluorescein 5-isothiocyanate is the 5-isomer of fluorescein isothiocyanate. Acts as a fluorescent probe capable of being conjugated to tissue and proteins; used as a label in fluorescent antibody staining procedures as well as protein- and amino acid-binding techniques.
Preparation
By 4-nitrophthalic anhydride and resorcinol at 195-200 ℃ for 10h, cyclized into a mixture of 3'-nitrofluorescein and 4'-nitrofluorescein, and then acetylated with acetic anhydride , add saturated sodium hydroxide-ethanol solution for hydrolysis to obtain 4'-nitrofluorescein. Then, 4'-nitrofluorescein was dissolved in ammonia water, and Nickel Reishi and hydrazine hydrate were added to react at room temperature for 2 hours, and then reduced to 4'-aminofluorescein. The most elevated by acylation, elimination of fluorescein isothiocyanate.
