Chameleon Labels for Staining and Quantifying Proteins

Wetzl, Bianca K.; Yarmoluk, S. M.; Craig, Douglas B.; Wolfbeis, Otto S.

doi:10.1002/anie.200460508

Cited by 111 publications

(105 citation statements)

References 21 publications

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“…The manufacturer's literature quotes excitation and emission maxima of 644 nm and 732 nm, respectively, for the unconjugated dye and 465 nm and 629 nm, respectively, for the reaction product of the reagent with a protein. This reagent appears to be identical to the Py-5 reagent introduced by Wolfbeis [11]. In particular, we consider the detection limits and dynamic range, and electrophoretic behavior in different separation buffers.…”

Section: Introductionmentioning

confidence: 99%

Reaction of fluorogenic reagents with proteins

Swearingen

Dickerson

Turner

et al. 2008

Journal of Chromatography A

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The fluorogenic reagent Chromeo P465 is considered for analysis of proteins by capillary electrophoresis with laser-induced fluorescence detection. The reagent was first used to label α-lactalbumin; the product was analyzed by capillary zone electrophoresis in a sub-micellar sodium dodecyl sulfate (SDS) buffer. The product generated a set of equally spaced but poorly resolved peaks that formed a broad envelope with a net mobility of 4 × 10 −4 cm 2 V −1 s −1 . The components of the envelope were presumably protein that had reacted with different numbers of labels. The mobility of these components decreased by roughly 1 % with the addition of each label. The signal increased linearly from 1.0 nM to 100 nM α-lactalbumin (r 2 = 0.99), with a 3σ detection limit of 70 pM. We then considered the separation of a mixture of ovalbumin, α-chymotrypsinogen A, and αlactalbumin labeled with Chromeo P465; unfortunately, baseline resolution was not achieved with a borax/SDS buffer. Better resolution was achieved with N-cyclohexyl-2-aminoethanesulfonic acid/ Tris/SDS/dextran capillary sieving electrophoresis; however, dye interactions with this buffer system produced a less than ideal blank.

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Section: Introductionmentioning

confidence: 99%

Reaction of fluorogenic reagents with proteins

Swearingen

Dickerson

Turner

et al. 2008

Journal of Chromatography A

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“…Labels L-1, 26 Py-1, 27 and Py-4 28 were synthesized as described in literature. Label Py-6 was synthesized by analogy to the protocol for Py-1 in a single reaction step from 1,1,3-trimethyl-1,3-dihydro-benz[e]indol-2-yliden)-acetaldehyde and 2,4,6-trimethylpyrylium tetrafluoroborate.…”

Section: Labelsmentioning

confidence: 99%

Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond Domain

Hoefelschweiger

Wolfbeis

2005

SLAS Discovery

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The authors demonstrate that the fluorescence lifetime of certain fluorescent labels is a useful parameter to detect affinity binding between biotin and streptavidin, as well as between biotinylated bovine serum albumin and streptavidin. The assay is performed in a microplate format, and lifetimes are determined using dye laser-induced fluorescence. Four fluorescent labels are presented that undergo a significant change in their lifetime upon affinity binding. The scheme, referred to as the fluorescence lifetime affinity assay, has several attractive features in that it requires single labeling only, represents a homogeneous assay, allows each of the 2 binding partners to be labeled, and is compatible with the standard microwell formats used in highthroughput screening. (Journal of Biomolecular Screening 2005:687-694)

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“…Finally, the fourth group of fusion molecules comprised either a SiFA 46−48 or DOTA moiety (for radiolabeling with 18 For 68 Ga) as well as the nearinfrared-emitting fluorescent dye, Py5. 49 These fusion molecules could, in principle, be applicable in in vivo PET imaging or combined PET/optical imaging of hD 2 receptors. From these conjugates, it should be possible to determine if the binding affinity of the fallypride moiety to the hD 2 receptor tolerates complex structure elements essential for radiolabeling and in vivo imaging such as SiFA, DOTA, and fluorescent dyes introduced within the carrier peptide part of the molecule.…”

Section: Journal Of Medicinal Chemistrymentioning

confidence: 99%

Shuttle–Cargo Fusion Molecules of Transport Peptides and the hD_2/3Receptor Antagonist Fallypride: A Feasible Approach To Preserve Ligand–Receptor Binding?

et al. 2014

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Access and use of this website and the material on it are subject to the Terms and Conditions set forth at Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. For the publisher's version, please access the DOI link below./ Pour consulter la version de l'éditeur, utilisez le lien DOI ci-dessous.http://doi.org/10.1021/jm5004123Journal of Medicinal Chemistry, 57, 10, pp. 4368-4381, 2014-04-30 Shuttle ABSTRACT: To determine if the conjugation of a small receptor ligand to a peptidic carrier to potentially facilitate transport across the blood−brain barrier (BBB) by "molecular Trojan horse" transcytosis is feasible, we synthesized several transport peptide−fallypride fusion molecules as model systems and determined their binding affinities to the hD 2 receptor. Although they were affected by conjugation, the binding affinities were found to be still in the nanomolar range (between 1.5 and 64.2 nM). In addition, homology modeling of the receptor and docking studies for the most potent compounds were performed, elucidating the binding modes of the fusion molecules and the structure elements contributing to the observed high receptor binding. Furthermore, no interaction between the hybrid compounds and P-gp, the main excretory transporter of the BBB, was found. From these results, it can be inferred that the approach to deliver small neuroreceptor ligands across the BBB by transport peptide carriers is feasible.

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Chameleon Labels for Staining and Quantifying Proteins

Cited by 111 publications

References 21 publications

Reaction of fluorogenic reagents with proteins

Reaction of fluorogenic reagents with proteins

Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond Domain

Shuttle–Cargo Fusion Molecules of Transport Peptides and the hD_2/3Receptor Antagonist Fallypride: A Feasible Approach To Preserve Ligand–Receptor Binding?

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Chameleon Labels for Staining and Quantifying Proteins

Cited by 111 publications

References 21 publications

Reaction of fluorogenic reagents with proteins

Reaction of fluorogenic reagents with proteins

Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond Domain

Shuttle–Cargo Fusion Molecules of Transport Peptides and the hD2/3Receptor Antagonist Fallypride: A Feasible Approach To Preserve Ligand–Receptor Binding?

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Product

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Shuttle–Cargo Fusion Molecules of Transport Peptides and the hD_2/3Receptor Antagonist Fallypride: A Feasible Approach To Preserve Ligand–Receptor Binding?