76823-03-5 Usage
Description
5-Carboxyfluorescein is a single isomer derivative of 5(6)-carboxyfluorescein that can be used to fluorescently label biomolecules through the interaction of carboxylic acid with primary amines. It demonstrates excitation/emission maxima of 492 and 518 nm, respectively.
Chemical Properties
Orange Solid
Uses
5-Carboxyfluorescein(5-FAM) contains a carboxylic acid that can be used to react with primary amines via carbodiimide activation of the carboxylic acid. It is a useful reagent for the preparation of hydrolytically stable fluorescent conjugates and is a useful starting material for the synthesis of other fluorescein-derives reagent. It has been principally used to develop a variety of green fluorescent reagents and small fluorescent molecules due to its relatively high absorbance characteristics, excellent fluorescence quantum yield, and good water solubility. The probe has also found application in studies of liposome-cell, cell-cell, and liposome-liposome interactions and in related studies on lipid bilayer structures.
Definition
ChEBI: 5-carboxyfluorescein is a monocarboxylic acid. It has a role as a fluorochrome. It is functionally related to a fluorescein (lactone form).
Biochem/physiol Actions
5-Carboxyfluorescein is used for the staining of cells to differentiate between live and dead cells. It helps in monitoring the integrity and intactness of the cell membrane. It is also used for the measurement of intracellular pH. 5-Carboxyfluorescein diacetate is a non-fluorescent molecule which is cleaved by intracellular esterases, thereby giving a green fluorescent product 5-carboxyfluorescein. In viable cells, the fluorescent molecule is impermeable to the cellular membrane.
Synthesis
5-Carboxyfluorescein is a mixture of 5- and 6-isomers prepared by the reaction of benzotricarboxylic anhydride with resorcinol and zinc chloride.Reaction: 1,2,4-Benzenetricarboxylic anhydride (also called 4-carboxyphthalic anhydride, 25.0 g, 0.13 mol) was added to a solution of 1,3-dihydroxybenzene (also called resorcinol, 28.6 g, 0.26 mol) in methane sulfonic acid (1M). An air condenser was attached to the flask and the reaction was heated at 85°C in an open vessel for 24 h. After cooling to room temperature, the reaction mixture was poured into 7 volumes of ice/water. An orange-yellow precipitate formed; this was collected by filtration and dried in an oven at 200°C. This residue was recrystallized two times from methanol/hexane to give 1.0 g of 6-carboxyfluorescein methanesulfonic acid adduct 2b. The mother liquors from this procedure were collected, the solvent was removed in vacuo, and the residues were recrystallized two times from ethanol/hexanes to give 3.2 g of 5-carboxyfluorescein methanesulfonic acid adduct 2a. The methane sulfonic esters 2 are easily converted to the 5- and 6-carboxyfluoresceins 1 by treatment with sodium hydroxide solution then neutralizing with aqueous HCl.
Check Digit Verification of cas no
The CAS Registry Mumber 76823-03-5 includes 8 digits separated into 3 groups by hyphens. The first part of the number,starting from the left, has 5 digits, 7,6,8,2 and 3 respectively; the second part has 2 digits, 0 and 3 respectively.
Calculate Digit Verification of CAS Registry Number 76823-03:
(7*7)+(6*6)+(5*8)+(4*2)+(3*3)+(2*0)+(1*3)=145
145 % 10 = 5
So 76823-03-5 is a valid CAS Registry Number.
InChI:InChI=1/C21H12O7/c22-11-2-5-15-17(8-11)27-18-9-12(23)3-6-16(18)21(15)14-4-1-10(19(24)25)7-13(14)20(26)28-21/h1-9,22-23H,(H,24,25)
76823-03-5Relevant articles and documents
The fluorescence signal for increasing the use of divalent metal
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Paragraph 0177; 0179, (2021/08/21)
Compositions comprising fluorescent or colored dyes and methods of using the same in combination with divalent metal salts to analyze samples are disclosed.
COMPOSITIONS COMPRISING A POLYMERIC DYE AND A CYCLODEXTRIN AND USES THEREOF
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Page/Page column 129, (2017/12/01)
Compositions comprising a polymeric dye and a cyclodextrin are provided. Methods associated with preparation and use of such compounds, for example in methods for detection of any analyte molecule, are also provided.
Site-specific, reversible and fluorescent immobilization of proteins on CrAsH-modified surfaces for microarray analytics
Schulte-Zweckel, Janine,Rosi, Federica,Sreenu, Domalapally,Schr?der, Hendrik,Niemeyer, Christof M.,Triola, Gemma
supporting information, p. 12761 - 12764 (2015/05/20)
A novel technique for protein immobilization onto CrAsH-modified surfaces is presented. This approach enables an efficient, reversible and fluorogenic immobilization of proteins. Moreover, expressed proteins can also be directly immobilized from cellular lysates without prior purification. The immobilized proteins are suitable for protein-protein interaction studies and the fluorescence enhancement upon immobilization can be employed for the direct detection of the immobilized protein without the need for secondary detection methods. This journal is
