Fluorophore label

To enhance understanding of macromolecular absorption via the lung we have performed a series of investigations on the transepithelial transfer of fluorophore labelled polyhydroxyethylaspartamides in the rat lung. Our results will be discussed... 

Table I. Transfer to Perfusate for Several Fluorophore-labelled Molecular Weight Distributions of Poly(hydroxyethylaspartamide)a...

The commercially available dicyanomethylene squaraine dye Seta-670-mono-NHS showed extremely low blinking effects and good photostability when used in single-molecule studies of multiple-fluorophore labeled antibodies [113]. Seta-670-mono-NHS and Seta-635-NH-mono-NHS were covalently labeled to antibodies and used in a surface-enhanced immunoassay [114]. From the fluorescence intensity and lifetime changes determined for a surface that had been coated with silver nanoparticles, both labeled compounds exhibited a 15- to 20-fold... 

Fluorophores and analytes derivatized with a fluorophoric label [56-105]... 

GPI-anchored proteins (GP-APs) are synthesized in the cytoplasm and their transport into the ER occurs during the process of acquiring a GPI anchor, which is ultimately sorted into the outer leaflet of plasma membranes [24], A FRET study of fluorophore-labeled GPI-APs in cultured... 

Fluorophore-labeled antibodies, 14 148 Fluorophores, commonly used, 14 148, 149t Fluoropolymers... 

Make it visible the fluorophore label can be visualized directly using fluorescent microscopy. The biotin label (see Sect. 6.2.1) can be detected using streptavidin conjugated with an enzyme the latter must be visualized through an enzyme chromogenic system. Incubate sections with an appropriate enzyme substrate until optimal color develops (see Sect. 2.3). 

When using an enzyme (peroxidase or alkaline phosphatase) marker, incubate sections in an enzyme substrate solution until desired stain intensity develops (see Sect. 2.3). A fluorophore label can be directly visualized in a fluorescent microscope. 

Primary antibodies blot excess blocking solution from sections and incubate for 60 min at room temperature or overnight at +4°C with a mixture of correspondingly diluted unlabeled primary antibodies raised in two different host species (e.g., mouse and rabbit). When using fluorophore-labeled primary antibodies as in direct immunostaining method (one antibody layer), you may skip step (6) with secondary antibodies for indirect immunostaining method (two antibodies layers). Wash sections in PBS for 3 x 3 min. 

When using biotin-labeled secondary antibody, you have first to visualize biotin with a fluorophore-labeled (strept)avidin employing ABC technique (see Sect. 6.2.1), before proceeding to counterstaining (step 7). 

Another approach to this persistent problem relies on haptenylation of primary antibodies. Hapten (e.g., biotin, digoxigenin or any fluorophore) can be covalently bound to the antibody via A-hydroxysuccinimide esters (NHS-ES) (see Sect. 2.1), or conjugated employing monovalent IgG Fc-specific Fab fragments (see Sect. 2.2). Haptenylated primary antibodies can be subsequently visualized with the use of secondary antibodies recognizing the corresponding hapten (Fig. 8.5). Fluorophore-labeled primary antibodies can be directly visualized in a fluorescent microscope. 

Antifade agents (free-radical scavengers) in aqueous mounting media prevent photobleaching of fluorophore labels under exposure to excitation light. 

IQ (iron quench) Pierce Biotechnology, Inc. Iron compound binds fluorophore-labeled peptides after phosphorylation and quenches fluorescence intensity... 

These enantioselective capillary columns showed extremely good performance in the CEC mode. Plate numbers in excess of 100,000 m could be easily achieved for a variety of amino acid derivatives (with chromophoric and fluorophoric labels) (Eigure 1.34a) as well as other chiral acids such as 2-aryloxycarboxylic acids. 

---