TRITC tetramethyl rhodamine

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. 

Fluorescence detection relies on the visualization of a secondary antibody that has been labeled with a fluorophore such as fluorescein (FITC), Texas Red, Tetramethyl rhodamine (TRITC), or R-phycoerythrin. Although this method of detection has a reduced sensitivity of twofold to fourfold compared to chemiluminescence detection, it presents a tenfold greater linear dynamic range, thus providing better linearity and better quantiflcation within the detection limits. Since secondary antibodies can be labeled with fluor-ophores of distinct colors, multiplexing (simultaneous detection of several antigens) of the same blot is feasible. 

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. 

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

BARS concentration by some 5-fold to 15-fold, based on the calculations that the intracellnlar concentration is around 20 //g/ml and on the assumption that 5-10% of the cell volume is injected. Prior to injection, the protein is mixed with 0.4 mg/ml fluorescein isothiocyanate (FITC)- or tetramethyl-rhodamine B isothiocyanate (TRITC)-labeled dextran (Molecnlar Probes) as a marker for the microinjected cells (Bonazzi et al., 2005). To give an example, in stndies of basolateral and apical transport (using the vesicular stomatitis virus glycoprotein and p75, respectively), the proteins were microinjected 1 h after the beginning of the 20° incubation in the transport assay. After injection, the cells were allowed to recover for 1 h before proceeding with the experimental protocol (Bonazzi et al, 2005). The BARS (p50-2) and dynamin (DYN2) antibodies were injected at 4.5 mg/ ml, 3 h before farther experimental procedures. 

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