27072-45-3

Basic information

• Product Name: Fluorescein isothiocyanate
• Synonyms: FluoresceinIsothiocyanate,Isomers1&11;Spiroisobenzofuran-1(3H),9-9Hxanthen-3-one, 3,6-dihydroxy-5(or 6)-isothiocyanato-;FITC, Fluorescein isothiocyanate;Fuorescein isothiocyanate;Fluorescein 5(6)-isothiocyanate,FITC, Fluorescein isothiocyanate;Fluorescein5(6)-isot;5(6)-FITC;Fluorescein 5(6)-isothiocyanat, mixture of isomers >90%
• CAS NO: 27072-45-3
• Molecular Formula: C₂₁H₁₁NO₅S
• Molecular Weight: 389.38
• EINECS: 222-042-0
• Mol File: 27072-45-3.mol
• Article Data: 7

Chemical Properties

• Melting Point: >360 °C(lit.)
• Boiling Point: 708.6°Cat760mmHg
• Flash Point: 382.4°C
• Appearance: /powder
• Density: 1.54g/cm³
• Vapor Pressure: 9.53E-21mmHg at 25°C
• Refractive Index: 1.753
• Storage Temp.: 2-8°C
• Solubility: DMSO (Slightly), Methanol (Slightly)
• Stability: Moisture Sensitive
• Merck: 14,4089
• BRN: 1407295
• CAS DataBase Reference: Fluorescein isothiocyanate(CAS DataBase Reference)
• NIST Chemistry Reference: Fluorescein isothiocyanate(27072-45-3)
• EPA Substance Registry System: Fluorescein isothiocyanate(27072-45-3)

Safety Data

• Hazard Codes: Xn
• Statements: 42/43-36/37/38-20/21/22-42
• Safety Statements: 22-36-26-24/25-45-36/37
• WGK Germany: 3
• F: 10
• Hazardous Substances Data: 27072-45-3(Hazardous Substances Data)

Usage

Description

Fluorescein isothiocyanate (FITC) is an amine-reactive derivative of fluorescein dye, characterized by its dark yellow powder form and hygroscopic nature. It is soluble in water and alkaline aqueous solutions, with an absorption maximum at 495 nm and an emission maximum at 525 nm. FITC is known for its strong green fluorescence, which disappears and precipitates when dissolved in an alkaline solution but reappears upon the addition of alkali. It is widely used as a label for antibodies and other probes in various applications, including fluorescence microscopy, flow cytometry, and immunofluorescence-based assays such as Western blotting and ELISA.

Uses

1. Used in Multifunctional Uniform Nanoparticles:
Fluorescein isothiocyanate is used as a fluorescent reagent in the development of multifunctional uniform nanoparticles composed of a magnetite nanocrystal core and a mesoporous silica shell. These nanoparticles are utilized for magnetic resonance, fluorescence imaging, and drug delivery applications.
2. Used in Fluorescent Labeling of Proteins:
FITC is employed as a fluorescent label for proteins, such as antibodies and lectins, in immunofluorescence applications. The isothiocyanate group reacts with amino terminal and primary amines in proteins, allowing for the attachment of a fluorescent label.
3. Used in Gel Chromatography:
Fluorescein isothiocyanate is used for protein labeling in gel chromatography, enabling the detection and analysis of proteins during the separation process.
4. Used as an Immunofluorescent Probe:
FITC is used to label antibodies for use as immunofluorescent probes, which are essential tools in the study of cellular and molecular processes.
5. Used in Hybridization Probes:
Fluorescein isothiocyanate is used to label oligonucleotides for hybridization probes, allowing for the detection and analysis of specific nucleic acid sequences.
6. Used in Detecting Proteins in Gels and on Western Blots:
FITC is utilized for detecting proteins in gels and on Western blots, providing a visual representation of protein expression and interactions.
7. Used as a Contact Sensitizer:
Fluorescein isothiocyanate isomer I has been proposed as a contact sensitizer, which may have potential applications in the study of allergic reactions and skin sensitization.

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.

InChI:InChI=1/C21H11NO5S/c23-12-2-5-16-18(8-12)26-19-9-13(24)3-6-17(19)21(16)15-4-1-11(22-10-28)7-14(15)20(25)27-21/h1-9,23-24H

Upstream product

• 1122-60-7 1-nitrocyclohexane 
• 5466-84-2 4-nitrophthalic anhydride 
• 75-15-0 carbon disulfide 
• 3326-34-9 fluoresceinamine 
• 463-71-8 thiophosgene 
• 14926-29-5 3',6'-bis(acetyloxy)-5-nitrospiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3-one 
• 108-46-3 recorcinol 
• 610-27-5 4-nitrophthalic acid 
• 3326-35-0 3',6'-dihydroxy-5-nitrospiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3-one 
• 2321-07-5 fluorescein 

Downstream Products

• 1394015-41-8 C₄₂H₃₄N₂O₉S 
• 1072920-58-1 C₄₈H₄₉N₃O₁₀S 
• 1047678-22-7 C₅₃H₆₉N₉O₁₇S 
• 1093758-49-6 C₄₃H₂₉NO₁₆S 
• 1023970-86-6 C₃₇H₃₄IN₃O₈S 

Relevant articles and documents

Preparation method of fluorescein probe with specific selectivity

The invention relates to the field of preparation of fluorescein probes with specific selectivity, and particularly discloses a preparation method of a fluorescein probe with specific selectivity. The method comprises the following steps: (1) mixing 4-nitrophthalic anhydride, resorcinol, nitrocyclohexane and anhydrous cobalt chloride, conducting heating and melting, adding zinc chloride, conducting reacting, then adding hydrochloric acid for reflux, and conducting filtering, washing and drying; (2) mixing a reducing solution containing Na2S and Na2SO3 with a product obtained in the step (1), conducting reacting, then dropwise adding glacial acetic acid until complete precipitation, and conducting filtering, washing and drying; and (3) mixing a mixed solvent containing methanol and triethylamine with a product obtained in the step (2), dropwise adding CS2, conducting reacting, dropwise adding a copper chloride aqueous solution, continuing reacting after dropwise adding, adjusting the pH value of the system to 1-2 by using hydrochloric acid, and carrying out reduced pressure distillation, washing, drying and column chromatography purification. According to the method, the conversion rate and the yield of the 5-fluorescein isothiocyanate are greatly improved.

Method for preparing 5-fluorescein isothiocyanate ester

The invention provides a method for preparing 5-fluorescein isothiocyanate ester. The method comprises the following steps: (1) reacting an original material 5-amino fluorescein with carbon disulfide in the presence of a catalyst and a solvent, so as to obtain fluorescein-5-amine disulfide formate; and (2) reacting fluorescein-5-amine disulfide formate, obtained in the step (1), in the presence of the catalyst and the solvent, so as to obtain 5-fluorescein isothiocyanate ester. The method has the beneficial effects that the technical operational safety is relatively high, the production cost is low, a synthetic process is simple, and the reaction condition is mild; and 5-fluorescein isothiocyanate ester is high in yield and wide in application range and can adequately meet the requirements of industrial production of products.

Fluorescent human EP3 receptor antagonists

Exchange of the lipophilc part of ortho-substituted cinnamic acid lead structures with different small molecule fluorophoric moieties via a dimethylene spacer resulted in hEP3R ligands with affinities in the nanomolar concentration range. Synthesized compounds emit fluorescence in the blue, green, and red range of light and have been tested concerning their potential as a pharmacological tool. hEP3Rs were visualized by confocal laser scanning microscopy on HT-29 cells, on murine kidney tissues, and on human brain tissues and functionally were characterized as antagonists on human platelets. Inhibition of PGE2 and collagen-induced platelet aggregation was measured after preincubation with novel hEP3R ligands. The pyryllium-labeled ligand 8 has been shown as one of the most promising structures, displaying a useful fluorescence and highly affine hEP3R antagonists.