Fluorescent assays

Figure 8. Simultaneous measurement of intracellular Ca and oxidant production in neutrophils. Cells were labeled with Quin-2 and suspended at 2 x lo cells/mL buffer. At time zero, 1 nJf FLPEP was added (upper trace in each panel). In addition, the receptor blocker tBOC was added (3 x 10" M) after 30 s to stop further binding of the stimulus (lower trace in each panel). The excitation wavelength was 3A0 nm. Top panel Quin-2 fluorescence determined on channel B (of Figure 1) using a Corion A90-nm interference filter. The crossover from the superoxide assay has been subtracted. Middle panel Oxidant production (superoxide equivalents) determined by the para-hydroxyphenylacetate assay. Fluorescence was observed at AOO nm (on channel A of Figure 1).

The analysis of cell-cycle progression was one of the earliest applications of flow cytometry (for review, see Darzynkiewicz et al., 2004). In this assay, fluorescence signals from cells stained with DNA-binding fluorochromes are plotted as DNA content histograms that may be analyzed by using histogram deconvolution software to quantify cell-cycle phase distributions (Rabinovitch 1994). Fluorochromes that are useful for this purpose are the plasma membrane-impermeant DNA stains, propidium iodide (PI),... 

Recent detection methods for glycan array include fluorescent assay, SPR, MALDI-TOF mass spectrometry, and nanoparticle assay. Fluorescence-based measurement is the prevalent principle for detecting binding to glycan microarrays. Rhodamine [9],... 

Labeled chemokines have been generated by a variety of procedures including fusion with alkaline phosphatase (1), derivatization with fluorescent probes (2), and radiolabeling (3,4). While homogeneous fluorescent based assays (fluorescence polarization, fluorescence energy transfer, etc.) will become... 

Protease is used to screen bead library for protease substrate positive beads in this fluorescent-quench assay fluoresce because the quencher is cleaved. 

Recently, a small molecule fluorophore phosphosensor technology referred as Pro-Q Diamond dye has been developed to detect and quantitate phosphorylated amino acids within peptides and proteins in microarrays. ° In addition to binding assays, fluorescence detection methods have also been developed for functional assays. For example, microarrays of quenched fluorescent substrates can be used to detect protease or esterase activities in the analytes. In this method, quenched fluorescent substrates are prepared by coupling the peptide substrate to coumarin, a fluorescent dye. These peptide substrates are then spotted onto the solid support... 

Key words Cardiotoxicity, In vitro assays for cardiotoxicity, Cardiac safety, Functional flux assays, Fluorescence-based assays, hERG, Patch clamp, Alternative tests, Microelectrode array, hESC-derived cardiomyocytes, FLIPER, Q-T interval, HEK 293, CHO, Embryonic stem cells, Differentiation, hiPSCs, Drug interaction, Safety pharmacology, ICH guidelines, Arrhythmia... 

For a number of assays, fluorescent-labeled analogues of a hgand are used. Some of those assays are of a quantitative nature, such as in fluorescence correlation spectroscopy (FCS), where the fluorescent-labeled analogue is used to determine binding kinetics (Fig. 5.5) [36]. FCS allows the direct detection of molecular interactions in solution. FCS monitors the random motion of a fluorescent molecule... 

Figure 6 Schematic depiction of a fluorescence anisotropy assay. Fluorescently tagged molecules are excited by plane-polarized light only molecules in the proper orientation are excited. Emitted light is detected in the original plane and in the perpendicular plane. The quantity of fluorescence observed in the two orientations is determined by the rate of tumbling, which depends on particle size and relates to binding.

Enzyme-linked immunosorbent assay/fluorescent enzyme immunoassay... 

In an immunoflorescence assay, fluorescent dyes such as fluorescein and rhodamine can be visualized on a microscope slide through the use of ultraviolet light or detected with instruments which measure their emissions. Fluorescent immunoassays are widely used in clinical laboratories. They are quite sensitive but may be time-consuming and reagents may be unstable and require special handling. 

Sample Handling. Enzymes Immobilized Enzymes Enzyme-Based Assays. Fluorescence Clinical and Drug Applications. Gas Chromatography Mass Spectrometry. Infrared Spectroscopy Near-Infrared. Isotope Dilution Analysis. Liquid Chromatography Column Technology Instrumentation. Sensors Overview. 

See also Chemiluminescence Overview. Chiroptical Analysis. Derivatizatlon of Analytes. Enzymes Enzyme-Based Assays. Fluorescence Overview Ciinicai and Drug Appiications. Gas Chromatography Coiumn Technoiogy Mass Spectrometry. Immunoassays Overview. Immunoassays, Applications Clinical. Immunoassays, Techniques Enzyme Immunoassays Luminescence Immunoassays. Infrared Spectroscopy Overview. Liquid Chromatography Column Technology Normal... 

See also Bioluminescence. Chemiluminescence Liquid-Phase. Electrophoresis Blotting Techniques. Enzymes Immobilized Enzymes Enzyme Assays. Fluorescence Quantitative Analysis. Forensic Sciences Blood Analysis. Immunoassays Overview. Immunoassays, Applications Clinical Forensic. Immunoassays, Techniques Radioimmunoassays. 

See also. Chemiluminescence Oven/iew. Derivatiza-tion of Analytes. Electrophoresis Oven/iew. Enzymes Oven/iew Immobiiized Enzymes Enzyme-Based Eiec-trodes Enzymes in Physioiogicai Sampies Industriai Products and Processes Enzyme-Based Assays. Fluorescence Clinical and Drug Applications. Immunoassays Overview Production of Antibodies. Immunoassays, Applications Clinical Food Forensic. Immunoassays, Techniques Radioimmunoassays Enzyme Immunoassays Luminescence Immunoassays. Mass Spectrometry Polymerase Chain Reaction Products. Nucleic Acids Chromatographic and Electrophoretic Methods Electrochemical Methods. Polymerase Chain Reaction. 

A bridge between natural and artificial macromolecular metal complexes is the interaction of metal ions/complexes with peptides/proteins [70], nucleic acids/DNA [71,72], enzymes [73], steroids [74], carbohydrates [75]. Biometal-organic chemistry concentrates on such complexes [15], The reason for the increasing interest in this field lays in medical applications of metal complexes [16,76] (cancer, photodynamic therapy of cancer -immuno-assays, fluorescence markers, enantioselective catalysis, template orientated synthesis of peptides) as exemplarily shown below. 

Labeled Lateral-flow assay fluorescent Nucleic acid for HPV 30 min 10 copies/pL for types 57... 

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