Fluorescent isothiocyanates

Hebert, G.A., Pittman, B., and Cherry, W.B. (1967) Factors affecting the degree of nonspecific staining given by fluorescent isothiocyanate labeled globulins./. Immunol. 98, 1204—1212. 

A further proof of principle was conducted by Cooney et al. who demonstrated the feasibility of the Andersen cascade impactor as a cell compatible deposition device [90], Permeability coefficients of fluorescent isothiocyanate-labeled dextrans after impaction as aerosols on Calu-3 cells were calculated. Deposition did not negatively affect the cell monolayer integrity. 

Peptide analysis, sequencing and synthesis All fluorescent dye labels with reactive groups for amine, dansyl chloride, dabsyl chloride. Fluorescent isothiocyanates. N-Terminal amino acid analysis, peptide sequencing, peptide synthesis, labeling peptides in solution, solid-phase synthesis of labeled peptides 37... 

Calibration. Many approaches have been used to calibrate flow cytometric measurements. Including the comparison of flow and nonflow techniques (radiolabels, spectrofluorometry). In recent years, commercial standards have been introduced which are calibrated in fluorescein equivalents/particle (e.g., 3,000 or 500,000). With labeled ligands, calibration requires determining the relative quantum yield of the ligand compared to pure fluorescein and using the standards to analyze the amount bound on cells. Our ligands (fluorescein isothiocyanate derivatives) are typically 50% as fluorescent as fluorescein. 

Labelling Na,K-ATPase with ATP analogues provides evidence for contribution from charged residues that are widely separated in the sequence of a subunit of Na,K-ATPase. The first indication came from ATP sensitive covalent insertion of fluorescein-isothiocyanate (FITC) into Lys ° in the a subunit [90], The strong fluorescence signal provides a convenient probe for monitoring conformational transitions in the proteins. Site-directed mutagenesis of Lys reduces the activity of... 

Abbreviations EPR, electron paramagnetic resonance FITC, fluorescein-5 -isothiocyanate lAEDANS, iV-iodoacetyl-N -(5-sulfo-l-naphthyl)ethylenediamine NCD, fluorescent yV-cyclohexyl-N -(4-dimethyl-amino-a-naphthyl)carbodiimide RITC, rhodamine-5 -isothiocyanate DPPE, dipalmitoylphosphatidyl-ethanolamine PE, egg phosphatidyl-ethanolamine ANS, 8-anilino-l-naphthalene sulfonate DPH, diphenylhexatriene e-ADP, l,iV -ethanoadenosine-5 -diphosphate TNP-ADP, 2 [3 ]-0-(2,4,6-trinitrophe-nyl)adenosine-5 -diphosphate. 

The nondestructive introduction of a fluorescent label would provide the molecule with a nonradioactive fluorophore, yet would preserve the option for direct radiolabelling of the fluorescent moiety with 125Iodine. This approach was pioneered by Nagasawa et al. (5) who reacted native or /V-desulfated heparins with a fluorescein isothiocyanate (FITC). The resulting degree of labelling was low... 

Muramoto K, Nokihara K, Ueda A, Kamiya H (1994) Gas-phase microsequencing of peptides and proteins with a fluorescent Edman-type reagent, fluorescein isothiocyanate. Biosci Biotechnol Biochem 58 300-304... 

Wu S, Dovichi NJ (1989) High-sensitivity fluorescence detector for fluorescein isothiocyanate derivatives of amino acids separated by capillary zone electrophoresis. J Chromatogr 480 141-155... 

Scheme 13 depicts the synthesis of compound 144 containing the isothiocyanate at both terminal positions prepared from the highly fluorescent tetramer 141 (Scheme 13) <2003JOC1512>. 

The isothiocyanate derivatives of DTT-4,4-dioxide 83 <2002CEJ5072> have been found to give chemically very stable fluorescent bioconjugates to be potentially used for fluorescent markers for biopolymers... 

Fluorescein isothiocyanate (FITC) is one of the most popular fluorescent probes ever created. An isothiocyanate derivative of fluorescein is synthesized by modification of its lower ring at... 

In the preparation of 15 nm core-shell fluorescent silica particles, Ow et al. (2004) reported that the naked core (2.2 nm) alone produced a fluorescence intensity of less than the free dye in solution, presumably due to dye quenching. However, upon addition of the outer silica shell around the core, the brightness of the particles increased to 30 times that of the free dye (using tetramethylrhodamine-5-(and 6)-isothiocyanate (TRITC)). They speculate that shell may protect the core from solvent effects, as evidenced by a lack of spectral shift upon changing the solvent in which the particles are suspended. 

In addition to the wide range of commercial probes, many other fluorescent molecules have been synthesized and described in the literature. Only a handful, however, are generally used to label antibody molecules. Perhaps the most common fluorescent tags with application to immunoglobulin assays are reflected in the main derivatives produced by the prominent antibody manufacturing companies. These include derivatives of cyanine dyes, fluorescein, rhod-amine, Texas red, aminomethylcoumarin (AMCA), and phycoerythrin. Figure 20.16 shows the reaction of fluorescein isothiocyanate (FITC), one of the most common fluorescent probes, with an antibody molecule. 

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