3301-79-9 Usage
Chemical Properties
Orange Solid
Uses
6-Carboxyfluorescein is derivative of fluorescein giving stable derivative upon conjugation with biopolymers.It has been used to check the plasma membrane integrity in sperm cells.
Preparation
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 oC 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 oC. This residue was recrystallized two times from methanol/hexane to give 1.0 g of 6- carboxyfluorescein methanesulfonic acid adduct preferentially crystallizes from MeOH/hexanes. 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 preferentially crystallizes from EtOH/hexanes. Finally, the mother liquors from this experiment were combined, evaporated to dryness, and recrystallized two times from methanol/hexanes to give another 3.0 g of 6-carboxyfluorescein methanesulfonic acid adduct preferentially crystallizes from MeOH/hexanes, making a total yield of 4.0 g, 40 %. Careful dropwise addition of conc. HCl(aq) to solutions of these methanesulfonic esters preferentially crystallizes from EtOH/hexanes and preferentially crystallizes from MeOH/hexanes in 4M sodium hydroxide gave 5-carboxyfluorescein and 6-carboxyfluorescein, respectively in near quantitative yield.
Definition
ChEBI: 6-carboxyfluorescein is a monocarboxylic acid. It derives from a fluorescein (lactone form).
Application
6-Carboxyfluorescein 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. Fluorescein is the most common fluorescent derivatization reagent for labeling biomolecules. In addition to its relatively high absorptivity, excellent fluorescence quantum yield, and good water solubility, fluorescein has an excitation maximum that closely matches the 488 nm spectral line of the argon-ion laser.
Biochem/physiol Actions
6-Carboxyfluorescein diacetate is a membrane permeable compound. It can be hydrolysed by intracellular esterases to form a membrane impermeable bright green fluorescent molecule, 6-carboxyfluorescein.
Check Digit Verification of cas no
The CAS Registry Mumber 3301-79-9 includes 7 digits separated into 3 groups by hyphens. The first part of the number,starting from the left, has 4 digits, 3,3,0 and 1 respectively; the second part has 2 digits, 7 and 9 respectively.
Calculate Digit Verification of CAS Registry Number 3301-79:
(6*3)+(5*3)+(4*0)+(3*1)+(2*7)+(1*9)=59
59 % 10 = 9
So 3301-79-9 is a valid CAS Registry Number.
InChI:InChI=1/C21H12O7/c22-11-2-5-14-17(8-11)27-18-9-12(23)3-6-15(18)21(14)16-7-10(19(24)25)1-4-13(16)20(26)28-21/h1-9,22-23H,(H,24,25)
3301-79-9Relevant articles and documents
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
, p. 12761 - 12764 (2014)
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
Synthesis and fluorescence properties of six fluorescein-nitroxide radical hybrid-compounds
Sato, Shingo,Endo, Susumu,Kurokawa, Yusuke,Yamaguchi, Masaki,Nagai, Akio,Ito, Tomohiro,Ogata, Tateaki
supporting information, p. 66 - 71 (2016/07/06)
Six fluorescein-nitroxide radical hybrid-compounds (2ab, 3ab, 4, and 5) were synthesized by the condensation of 5- or 6-carboxy-fluorescein and 4-amino-TEMPO (2ab), 5- or 6-aminofluorescein and 4-carboxy-TEMPO (3ab), and fluorescein and 4-carboxy-TEMPO (4), or by reaction of the 3-hydroxyl group of fluorescein with DPROXYL-3-ylmethyl methanesulfonate (5). Fluorescence intensities (around 520 nm) after reduction of the radical increased to 1.43-, 1.38-, and 1.61-folds for 2a, 2b and 3b respectively; 3a alone exhibited a decrease in intensity on reduction. Since 4 was readily solvolyzed in PBS or even methanol to afford fluorescein and 4-carboxy-TEMPO, its fluorescence change could not be measured. Hybrid compound 5 containing an ether-linkage between the fluorescein phenol and 3-hydroxymethyl-DPROXYL hydroxyl centers, was stable and on reduction, showed a maximum increase (3.21-fold) in relative fluorescence intensity in PBS (pH 5.0), despite its remarkably low absolute fluorescence intensity.
Design, synthesis, and characterization of fluorescent cobalamin analogues with high quantum efficiencies
Lee, Manfai,Grissom, Charles B.
supporting information; experimental part, p. 2499 - 2502 (2009/10/24)
Cobalamin tethered to fluorescein or Rhodamine 6G has been synthesized and characterized. The fluorophore is conjugated to the ribose-5-OH of cobalamin through a rigid linker to prevent the fluorophore from folding back through space and interacting with
