Fluorescence detection protein labeling

Oswald B, Gruber M, Bohmer M, Lehmann F, Probst M, Wolfbeis OS (2001) Novel diode laser-compatible fluorophores and their application to single molecule detection, protein labeling and fluorescence resonance energy transfer immunoassay. Photochem Photobiol 74 237-245... 

Extremely high dynamic range and exquisite sensitivity is produced by laser-induced fluorescence of FQ-labeled proteins. The dynamic range exceeds 250,000, and the detection limit is in the high yoctomole range for FQ-labeled proteins. 

Yan W, Sloat AL, Yagi S, Nakazumi H, Colyer CL (2006) Protein labeling with red squarylium dyes for analysis by capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis 27 1347-1354... 

Oswald B, Patsenker L, Duschl J, Szmacinski H, Wolfbeis OS, Terpetschnig E (1999) Synthesis, spectral properties, and detection limits of reactive squaraine dyes, a new class of diode laser compatible fluorescent protein labels. Bioconjugate Chem 10 925-931... 

When cells are suspended in a biological fluid or culture medium, both serum proteins and cells interact with the surface substrate. Serum protein adsorption behavior on SAMs has been examined with various analytical methods, including SPR [58-61], ellipsometry [13, 62, 63], and quartz QCM [64—66]. These methods allow in situ, highly sensitive detection of protein adsorption without any fluorescence or radioisotope labeling. SPR and QCM are compatible with SAMs that comprise alkanethiols. In our laboratory, we employed SPR to monitor protein adsorption on SAMs. 

The nonreactive base structures of cyanine dyes (or carbocyanines) have been used for many years as components in photographic emulsions to increase the range and sensitivity of film and also in CD-R and DVD-R optical disks to record digital information. A major innovation came when Ernst et al. (1989) and Waggoner et al. (1993) recognized that cyanine dyes would make excellent labels for fluorescence detection, and for this reason, they synthesized reactive dye derivatives, which then could be covalently attached to proteins and other molecules. 

A.D. Presley, K.M. Fuller and E. A. Arriaga, MitoTracker Green labeling of mitochondrial proteins and their subsequent analysis by capillary electrophoresis with laser-induced fluorescence detection. J. Chromatogr.B, 793 (2003) 141-150. 

Examples of its use include protein and nucleic acid detection, enzyme-labelled fluorescence, in the intrinsic fluorescence of normal and cancer cells, as external... 

Fig. 3. Detection of a synthesized protein by fluorescent labeling. Cell-free protein synthesis was carried out with or without the use of mRNA transcribed from a linearized expression done containing the p-gaiactosidase gene, and the synthesized protein was labeled by FluoroTect. The translational reaction mixtures were resolved by 12.5% SDS-PAGE. Detection of labeled protein was performed using a laser-based fluorescent scanner (FX pro, Bio-Rad, Hercules, CA). Lanes 1 and 2 represent negative control (absence of mRNA) and p-galactosidase, respectively.

Following procedures are performed between the lEF (step 4) and equilibration procedure (step 5) for the simultaneous protein labeling and detection. A fluorescence labeling is performed between the lEF and equilibration procedures (In-between staining). 

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