Fluorescendy labeled

Sample (target) preparation The sample (RNA, cDNA) is fluorescendy labeled. 

Purify the fluorescendy labeled glycoprotein(s) by gel filtration using a Sephadex G-25 column. 

A general protocol for the use of DCIA for fluorescendy labeling proteins that contain sulfhydryl residues may be obtained by following the method discussed for AMCA-HPDP (previous section). After purification of the labeled protein, the F/P ratio of fluorophore incorporation may be determined by measuring its 382/280 nm absorbance ratio. 

In some cases, the preparation of a fluorescendy labeled antibody is not even necessary. Particularly, if indirect methods are used to detect antibody binding to antigen, then preparing a fluorescendy labeled primary antibody is not needed. Instead, the selection from a commercial source of a labeled secondary antibody having specificity for the species and class of primary antibody to be used is all that is required. However, if the primary antibody needs to be labeled and it is not manufactured commercially, then a custom labeling procedure will have to be done. 

Agonist binding to receptors on intact cells has been studied by investigating FRET between a fluorescendy labeled agonist and receptors... 

An effective approach to study tRNA dynamics that is amenable to measurements of changes in both kinetics and amplitude is by fluorescence labeling at a site-specific position as a probe for local environment changes, either within the tRNA molecule or in the contact with a binding partner. The clear advantage of the fluorescence approach is the ability to monitor environmental changes continuously and in real time. Such fluorescendy labeled tRNAs were first used to study tRNA reactions on the ribosome... 

Competitive Format. Although UV detection can be used in the competitive format, its sensitivity is not high enough to provide a necessary limit of detection (LOD). Therefore, in almost all cases reported in the literature so far, LIF detection is used and, consequently, one of the reagents is fluorescendy labeled. 

As in the competitive mode of conventional immunoassays, at least one of the reagents must be limited to provide competition. If the analyte of interest is Ag, then a known amount of fluorescendy labeled Ag (so-called fluorescence tracer) is added to the sample together with a limited amount of antibody Ab. Analyte, present in a sample, competes with the tracer for a limited number of antibody-binding sites. The affinity reaction which occurs in such a mode is described by... 

The idea of using CIEF to separate bound from free antibody was realized for the first time by Shimura et al. [25], These authors quantified human growth hormone (hGH) using tetramethylrodamine/iodoacetamide-labeled anti-hGH Fab fragment. Excess fluorescendy labeled Fab was added to solutions with variable amounts of hGH and, upon incubation, Fab -Ag complex was separated from the excess Fab by CIEF. LIF detection was used, which provided a very low detection limit of 5 x 10 12 M of methionyl hGH. In this report different forms of Ag were detected simultaneously in one run, since complexes of labeled Fab with non-, mono-, and di-deaminated variants of met-hGH exhibited different isoelectric points and hence could be resolved by CIEF. 

The binding is visualized by using fluorescendy labeled molecules and scanning the chip with a computer. The pattern of fluorescent labels then indicates which mRNA or proteins are being expressed in the samples. 

The construction of fluorescendy labeled receptors uses standard molecular biology techniques. The fluorescent probe must be inserted in the C-terminal region to conserve the original signal peptide of the receptor. The fusion involves elimination of the receptor stop codon and insertion of the receptor into the fluorescent protein vector, separated by several additional amino acids. Transfect cells and analyze for correct expression and function. 

Dissociation constants can be estimated using affinity capillary electrophoresis (ACE)." In these experiments, a constant concentration of the nucleic acid pool is titrated with increasing concentrations of target. Use of fluorescendy labeled sequences is recommended since LIE detection... 

J.D. (2002) Reagendess pH-based biosensing using a fluorescendy-labelled dextran co-entrapped with a hydrolytic enzyme in sol-gel derived nanocomposite films. Anal. Chim. Acta, 4S17,47 59 (i) Frenkel-Mullerad, H. and Avnir, D. (2005) Sol-gel materials as efficient enzyme protectors preserving the activity of phosphatases under extreme pH conditions. /. Am. Chem. Soc., 127 (22), 8077-8081 (j) Frenkel-Mullerad, H.,... 

Saleh, S. M. Mueller, R. Mader, H. S. Duerkop, A. Wolfbeis, O. S. Novel multicolor fluorescendy labeled silica nanoparticles for interface fluorescence resonance energy transfer to and from labeled avidin. Anal. Bioanal. Chem. 2010,398,1615-1623. 

State, respectively. These values are in good agreement with the obtained by DLS, which were 265 and 168 nm. When a fluorescendy labeled biopolymer was incorporated into the multilayer system, single or... 

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