3326-32-7 Usage
Description
Fluorescein 5-isothiocyanate (5-FITC) is an isomer of fluorescein isothiocyanate (FITC), which is commonly used as a mixture of the 5- and 6-isothiocyanate isomers. It reacts with amine and thiol groups to form conjugates with proteins, lipids, and other molecules for detection by a variety of fluorescent-based applications. 5-FITC displays excitation/emission maxima of 494/519 nm, respectively.
Chemical Properties
Orange-yellow cryst.
Uses
Different sources of media describe the Uses of 3326-32-7 differently. You can refer to the following data:
1. Fluorescein -5-isothiocyanate is a kind of fluorescent marker for biochemical applications. Reacts under mild conditions with primary amines and it is also used in modification of actin at lys-61, labelling of FAB and the modification of thiol groups.
2. Fluorescein isothiocyanate isomer I has been used for the staining of conidia.
3. Fluorescein isothiocyanate is used as fluorescent labeling reagent for proteins and for rapid identification of pathogens in fluorescent antibody technique. It is also used as a reagent for microsequencing of proteins and peptides (HPLC). On celite it is used for the labeling of fibrinogen.
4. Fluorescent labeling reagent for proteins. Used in the fluorescent antibody technique for rapid identification of pathogens
Definition
ChEBI: 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.
General Description
In immuno histology/ cytology, Fluorescein-5-isothiocyanate (FITC) for biochemistry and for fluorescence microscopy, serve as marker substance for the visualization and for the detection of tissue-bound bio molecules like e.g. antigens, lectins, various proteins, peptides, nucleic acids, oligo- and polysaccharides in samples of human origin. FITC-marked antibodies bind specifically to the appropriate target structures in the tissue. An optimum FITC-protein (antibody) ratio is important for a good result.FITC is an IVD registered product and CE certified, thus can be used for clinical diagnostic purposes. For more details, please see instructions for use (IFU). The IFU can be downloaded from this webpage.
Purification Methods
It is made from the pure 5-amino isomer. Purify it by dissolving it in boiling Me2CO, filtering and adding pet ether (b 60-70o) until it becomes turbid. If an oil separates, then decant it and add more pet ether to the supernatant and cool. Orange-yellow crystals separate, collect and dry them in vacuo. It should give one spot on TLC (silica gel) in EtOAc/pyridine/AcOH (50:1:1) and in Me2NCHO/CHCl3/28% NH4OH (10:5:4). IR (Me2SO): 2110 (NCS) and 1760 (C=O) cm-1. The max 1HNMR spectra in Me2CO-d6 of the 5-and 6-isomers are distinctly different for the protons in the *benzene ring; the UV in phosphate buffer pH 8.0 shows a at ~490nm. [Sinsheimer et al. Anal Biochem 57 2271974, McKinney et al. Anal Biochem 7 74 1964, Beilstein 19 III/IV 4337.]
Check Digit Verification of cas no
The CAS Registry Mumber 3326-32-7 includes 7 digits separated into 3 groups by hyphens. The first part of the number,starting from the left, has 4 digits, 3,3,2 and 6 respectively; the second part has 2 digits, 3 and 2 respectively.
Calculate Digit Verification of CAS Registry Number 3326-32:
(6*3)+(5*3)+(4*2)+(3*6)+(2*3)+(1*2)=67
67 % 10 = 7
So 3326-32-7 is a valid CAS Registry Number.
InChI:InChI=1/C21H11NO5S/c23-12-2-5-15-18(8-12)27-19-9-13(24)3-6-16(19)20(15)14-4-1-11(22-10-28)7-17(14)21(25)26/h1-9,23H,(H,25,26)
3326-32-7Relevant articles and documents
Preparation method of fluorescein probe with specific selectivity
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, (2021/05/29)
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
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Paragraph 0044; 0045; 0046; 0047, (2017/07/19)
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
Tomasch, Miriam,Schwed, J. Stephan,Kuczka, Karina,Meyer Dos Santos, Sascha,Harder, Sebastian,Nuesing, Rolf M.,Paulke, Alexander,Stark, Holger
supporting information, p. 774 - 779 (2012/11/13)
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.
