Rhodamine isothiocyanate

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. 

Antibodies were labeled with an extrinsic fluorescent probe for the generation of a fluorescent signal during biosensor detection. The fluorescent probe used was tretramethyl-rhodamine-5-isothiocyanate (TRITC) purchased from Molecular Probes, Inc., Eugene, Oregon and stored in a desiccator at less than 0 °C prior to use. 

Abbreviations AMCA, 7-amino-4-methylcoumarin B-PE, B phycoerythrin Cy, cyanine DAMC, diethylaminocoumarin FITC, fluorescein isothiocyanate RB-200-SC, lissamine rhodamine sulfonylchloride R-PE, R phycoerythrin SITS, 4-acetamido-4 -isothiocyanato-stilbene-2,2 -disulfonic acid TRITC, tetramethyl rhodamine isothiocyanate XRITC, rhodamine X isothiocyanate. Information obtained from refs. 2, 9, and 10. 

The second strategy uses combinations of different antibodies coupled to fluorochromes with distinct emission maxima (5,9). The most relevant fluoro-chromes for combined antigen detection are fluorescein isothiocyanate (FITC abs. max. 494 nm, emiss. max. 517 nm), rhodamine isothiocyanate (TRITC ... 

The most used FIA reagents conjugate a fluorophore such as fluorescein—isothiocyanate (FITC) or rhodamine—isothiocyanate to antibody (or antigen) free amino groups. Examples of other commonly used fluorophores for FIA and their spectral characteristics are presented in Table 3. FIA assays are available in sandwich and competitive formats similar to EIAs. Unlike EIA kits which can be used directly with visual color determination, FIAs require a fluorometer, and thus are primarily laboratory-based. 

Gottschlich et al. [26] described the separation of tetramethyl rhodamine isothiocyanate (TRITC)-labeled tryptic peptides of (3-casein. The field strength was 220 V/cm in the NCEC channel with lOmM sodium borate with 30% (v/v) acetonitrile as mobile phase. Throckmorton et al. [27] described the separation of papain inhibitor, proctolin, opioid peptide (a-casein fragment 90-95), Ile-angiotensin III and angiotensin III on a porous polymer monolith... 

Fluorescein isothiocyanate (FITC) or tetramethyl-rhodamine isothiocyanate (TRITC). 

The red-emitting rhodamine derivatives are constructed around the same basic xanthene framework as is fluorescein (2). Tetramethyl-rhodamine isothiocyanate (TRITC) has been widely employed for immunofluorescence. Additional derivatives of rhodamine available for conjugation to antibodies include lissamine rhodamine sulfonyl chloride (RB-200-SC), rhodamine B isothiocyanate (RBITC), rhodamine X isothiocyanate (XRITC), and Texas Red (Molecular Probes, Inc.). The spectra of XRITC and Texas Red are shifted to longer wavelengths compared to those of other rhodamines, which makes them particularly useful for combination with fluorescein in dual-labeling procedures see Section 5, below). Of the two, Texas Red, which is more hydrophilic and less likely to precipitate proteins upon conjugation (12), is more commonly employed. 

Fluorescence resonance energy transfer (FRET) is a technique that has been used to measure distances between pairs of proximal fluorochromes. A suitable pair consists of a donor fluorochrome, which has an emission spectrum that significantly overlaps with the absorption spectrum of an acceptor fluorochrome (2). With the availability of monoclonal antibodies to many cell-surface determinants, intramolecular distances between nearby epitopes and intermolecular distances between adjacent cell-surface macromolecules can be investigated to analyze molecular interactions influencing important cellular events. Such monoclonal antibodies can be conjugated to fluorescein-isothiocyanate (FITC) as the donor, and either tetramethyl-rhodamine-isothiocyanate (TRITC) or phycoerythrin (PE) as the acceptor. 

Fluorescent labelling of the particles for confocal microscopy may be achieved in three ways. The first involves the use of polymerisable dyes. These dyes have been chemically modified to include a reactive group that can be chemically attached to the particle as they are produced. The advantage of this procedure is that the dye will not leave the particle once it is incorporated. For sterically-stabilised PMMA particles this involves adding a methacrylate group to the dye, and several such procedures [23-27] have been described in the literature. The most commonly used dye is 7-nitrobenzo-2-oxa-l,3-diazole-methyl methacrylate (NBD-MMA) [26,27], which is excited at 488 nm and emits at 525 nm, while the red end of the spectrum is well served by (rhodamine isothiocyanate)-aminostyrene (RAS) [27]. 

Finally, a double labelling with antibodies and a viability substrate can be performed. De Vos and Nelis (2003, 2006) combined ChemChrome V6 with tetram-ethyl rhodamin isothiocyanate (TRITC) labelled antibodies for the detection of Aspergillus fumigatus. In these approaches, the ChemChrome reagent, yielding green fluorescence, ensures the primary detection by the ChemScan, whereas the TRITC label results in red fluorescence, to be observed microscopically. 

Also aiming at biomedical applications are nanoscaled hydrogels, prepared in inverse miniemulsion. In crosslinked poly(oligo(ethylene glycol) monomethyl ether methacrylate) (POEOMA) nanogels hydrophilic dyes as the polymeric dye (rhodamine isothiocyanate (RITC) dextran) [41], rhodamine in combination with the drug doxorubicin [42] or gold nanoparticles with bovine serum albumin [43] could be encapsulated. 

Figure 41 shows the XPS spectra for the N Is region of adsorbed (Type I) and chemically bound (Type II) samples of rhodamine isothiocyanate as well as rhodamine B. Clear differences between them are observed curve a is from a type II sample and is centered around 399.5 eV curves b and c show increasing shifts toward larger binding energies. They correspond respectively to a type I sample (physically adsorbed dye from aqueous solutions), subsequently washed with water, and to rhodamine B type I sample from an ethanolic solution. 

Tetramethyl rhodamine Isothiocyanate (TRITC) [95197-95-8] 529 (492) 596 (518) Protein fluorochrome, used for immunofluorescence... 

Fig. 14 (a) Gross examination of subdermally implanted PDI-sucrose (white arrow) exhibiting absence of tissue necrosis, redness, and edema around the polymer 6 weeks after implantation. Green arrows indicate capillaries growing towards the polymers. Confocal microscopic examination of vascularization (white arrows) in the (b) superficial layers and (c) deeper layers of subdermally implanted polymer after 3 weeks. The polymer is seen as green because of autofluorescence and capillaries are stained red because of the intravenous injection of 0.2 pm of fluorospheres containing the fluorescent red dye rhodamine isothiocyanate [170]... 

ABEI, M(4-ami nobutyl )-Methylisolu mi nol BSA, bovine serum albumin CL, chemiluminescence DNPO, tas-(2,4-dinitrophenyl)oxalate ECL, electrogenerated chemiluminescence EMMA, electrophoretically mediated microanalysis EY, eosine Y FR, lluorescamine HRP, horseradish peroxidase ILITC, isoluminol isothiocyanate LOD, limit of detection RITC, rhodamine B isothiocyanate TCPO, Mv-(2,4,6-trichlorophenyl)oxalate TEA, triethylamine TRITC, tetramethylrhodamine isothiocyanate. 

Consequently, the research work of Hara s group continued focusing on the improvement of protein determination using CE combined with online CL detection. By replacing EY by the Rhodamine B isothiocyanate (RITC) dye in the binary complexes formed with the proteins BSA or human serum albumin (HSA) and using a different imidazole buffer solution of pH 6, the sensitivity was increased [72], However, best detection limits for these determinations were found employing the tetramethylrhodamine isothiocyanate isomer (TRITC) dye, left for 4 h with a standard solution of BSA in acetonitrile followed by introduction into the capillary. For BSA, a detection limit of 6 nM was reached [73],... 

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