Labeling with fluorescein

FIG. 9 Confocal laser scanning micrograph of a hollow polymer capsule. The polymer capsule was obtained from polymer multilayer-templated FDA microcrystals after removal of the colloidal core. The FDA microcrystals were coated with SDS and 11 polyelectrolyte layers [(PAH/PSS)3/PAH/ (PSS/PAH-FITC)2]. (PAH-FITC = PAH labeled with fluorescein isothiocyanate.) The microcrystal core was removed by exposure of the coated microcrystals to ethanol, causing solubilization of FDA. 

The azide can also react with phosphine derivatives through the Staudinger ligation. Azidophenylalanine was incorporated into the Z-domain protein in E. coli or into peptides displayed on phage, and was labeled with fluorescein-derived phosphines in phosphate buffer at room temperature (Figure 9(b)). ... 

In a more recent report, Kabisch et al. (1999) noted the purification of PR, also from C. sticklandii. The purified enzyme contained three subunits (SDS-PAGE), two of which were labeled with fluorescein thiosemicar-... 

A limitation of the flow cytometric binding assay has been the precise determination of the receptor affinity and calculation of the receptors per cell. This limitation appears to have been overcome by the development of fluorescein and phycoerythrin compensation-calibration standards (Flow Cytometry Standards Corp., Research Triangle Park, NC). These standards have made it possible to quantify the fluorescence intensity of samples labeled with fluorescein or phycoerythrin, and relate the intensity to molecules of equivalent soluble fluorochrome. These standards have been utilized in quantitative studies of neutrophil chemoattractant-ligand interaction (4). 

Abnova FISH probes and Dako FISH probes are labeled with fluorescein isothiocyanate (FITC) and Texas Red (sulforhod-amine lOI acid chloride) haptens, and the probe hybridization sites can be visualized with a BISH detection kit including anti-FITC and anti-Texas Red antibodies. 

Schmerr and Jenny established a CE-based immunoassay for the detection of prion protein (24). In this competitive assay, peptides derived from the prion protein and labeled with fluorescein were used. This allowed them to distinguish scrapie-infected brain preparations from noninfected. For identification, the ratio between the peaks resulting from the free and the com-plexed peptide with a specific antibody was used. The results were in agreement with other data on the brain preparations achieved by Western blot analysis. The CE-based assay provides the advantage of direct detection of the scrapie protein in blood and tissue preparations with high sensitivity. Furthermore, due to the small sample amount needed for analysis, the CE-based assay is applicable to the putative diagnostics of prion protein in body fluids. 

Figure 7. Elution profile for a sample of myoglobin which was partially cleaved with cyanogen bromide and labeled with fluorescein isothiocyanate. Absorbance at 280 fim, (broken line) and fluorescein fluorescence, 493-nm excitation, and 530-nm emission, (solid line) are plotted vs. fraction number. The identities of components A, B, C, D, and E were inferred on the basis of their Mav s (see...

When unlabelled 5 -primer is used for DNA amplification, the PCR products are labelled with fluorescein following the protocol explained in next section (Section 37.4.2). 

Figure 11.2 Subcellular distribution of fluorescein-labeled DNA fragments and plasmid DNA following microinjection into the cytoplasm. Double stranded circular plasmid DNA (3 kb and 6 kb) and DNA fragments (20,100,250 and 1 kb) were covalently labeled with fluorescein and microinjected into the cytoplasm of adherent HeLa cells as described in Lukacs et al., 2000. Following micro injection, cells were either fixed or incubated for 45 minutes at 37 °C and the distribution of DNA was visualized by fluorescence microscopy, (see Color Plate 10)...

In order to observe the behavior of chitosan-coated liposomes more precisely, chitosan was labeled with fluorescein isothiocyanate (FITC) via chemical reaction at the isothiocyanate group of FITC and the primary amino group of chitosan the liposomes (Lips) were marked by incorporation of Dil into the liposomal formulation. FITC-labeled chitosan (FITC-CS), non-coated liposomes, and FITC-labeled chitosan-coated liposomes (FITC-CS-Lips) were intragastrically administered into male Wistar rats, and then the behavior of the molecules was visualized by CLSM (Thongborisute et al. 2006a). The results demonstrated that the chitosan molecules themselves, as well as the liposomes, could penetrate across the intestinal mucosa. Moreover, the CLSM images demonstrated a lack of separation of the chitosan molecules from the surface of the liposomes after the administration of chitosan-coated liposomes. 

Sassaroli, M., Bucci, E., Liesegang, J., Fronticelli, C. and Steiner, R.F. (1984). Specialized functional domains in hemoglobin dimensions in solution of the apohemoglobin dimer labeled with fluorescein iodoacetamide. Biochemistry, 23,2487-2491. 

Peptide labeled with fluorescein and coumarin is quenched until cleavage by protease, modification by phosphorylation, or dephosphorylation by kinase or phosphatase produces resistance to proteolytic cleavage... 

Oppenheim RC, Stewart NF (1979) The manufacture and tumor cell uptake of nanoparticles labelled with fluorescein isothiocyanate. Drug Dev Ind Pharm 5(6) 563-572... 

The FRAP method has been applied to the measurements of molecular lateral diffusion of molecules adsorbed at the interface of equilibrium common thin foam films and of black foam films [39-43], Initially Clark et al. reported FRAP measurement of surface diffusion of the fluorescence probe 5-N(octadecanoyl)aminofluorescein incorporated into foam films stabilised with NaDoS [39]. Then followed the measurements of protein-stabilised foam films where the protein was covalently labelled with fluorescein [40,41], Studies of FRAP measurements of surface lateral diffusion in equilibrium phospholipid common thin foam films and black foam films were also reported [42,43]. 

Fluorescence microphotolysis can be used to measure diffusion in single cellsand nuclear envelope permeability. Pyranine probes have been used to measure internal pH changes in Escherichia coli membrane vesicles. Probe measurements of intralysosomal pH in living cells and perturbation of pH has been described by Ohkuma and Poole.Fluorescence polarization of six membrane probes in embryonal carcinoma cells have been measured by a cell sorter,Fluorescence of pancreatic islets labelled with fluorescein diacetate has been used to show the effects of cations, ionophores, and hypoglycaemic sulphonylureas. ... 

Either, primary andbody labeled with fluorescein or primary andbody (mouse monoclonal) and sheep andmouse Ig, fluorescein-labeled. 

One of the earliest efforts of qualitative measurement of a protein (human serum albumin) in a microchip-based device was based on bead agglutination in a microchamber (approximately lOjaL). Subsequently, several quantitative immunoassays have been performed using microchip electrophoretic systems that permit separation and quantitation of free- and bound-labeled antigens in competitive assays (see Chapter 5). Most are carried out in channels micro-machined into fused silica substrates. Early work on quantitative assays achieved measurement of cortisol in serum.The assay used cortisol labeled with fluorescein and an argon laser detector at 488 nm and required only 80 pL of a 40x dilution of serum as the sample. Other capillary electrophoresis-based assays for a variety of antibodies have also been developed that include immunoglobulins (IgG, IgA, and IgM), antibovine serum albumin, and antiestradiol. ... 

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