High Throughput Preparation of Peptide Arrays Containing Two Fluorescent Dyes Focusing on Practical Protein Detection Systems

2010 ◽  
pp. 738-739
Author(s):  
Kiyoshi Nokihara ◽  
Takafumi Ohyama ◽  
Koichi Yonemura ◽  
Yasuo Oka ◽  
Kenji Usui ◽  
...  
Keyword(s):  
High Throughput ◽  
Fluorescent Dyes ◽  
Protein Detection ◽  
Peptide Arrays ◽  
Detection Systems

scholarly journals Novel Signal Amplification Technology with Applications in DNA and Protein Detection Systems

BioTechniques ◽  
2001 ◽  
Vol 30 (6) ◽  
pp. 1268-1272
Author(s):  
C.M. Lisle ◽  
S. Bortolin ◽  
A.S. Benight ◽  
R.A. Janeczko ◽  
R.L. Zastawny
Keyword(s):  
Signal Amplification ◽  
Protein Detection ◽  
Detection Systems

scholarly journals Novel Electrochemiluminescent Assays for Drug Discovery

2000 ◽  
Vol 5 (1) ◽  
pp. 45-48 ◽  
Author(s):  
Maura C. Kibbey ◽  
David MacAllan ◽  
James W. Karaszkiewicz
Keyword(s):  
Drug Discovery ◽  
High Precision ◽  
High Throughput ◽  
High Performance ◽  
Dynamic Range ◽  
Wide Dynamic Range ◽  
Receptor Ligand ◽  
Enzymatic Assays ◽  
Detection Systems ◽  
Protein Assays

IGEN's ORIGEN® technology, which is based on electrochemiluminescence, has been adopted by a number of research and bioanalytical laboratories who have recognized its exquisite sensitivity, high precision, wide dynamic range, and flexibility in formatting a wide variety of applications. IGEN's M-SERIES™ marks the introduction of the second generation of detection systems employing the ORIGEN technology specifically repackaged to address the needs of the high throughput laboratories involved in drug discovery. Assays are formatted without wash steps. Users realize the high performance of a heterogeneous technology with the convenience of a homogeneous format. The M-SERIES platform can address enzymatic assays (kinases, proteases, helicases, etc.), receptor-ligand or protein-protein assays, immunoassays, quantitation of nucleic acids, as well as other applications. Recent assay formats will be explored in detail.

High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

ACS Nano ◽  
2022 ◽  
Author(s):  
Connie Wu ◽  
Tyler J. Dougan ◽  
David R. Walt
Keyword(s):  
High Throughput ◽  
Protein Detection

scholarly journals High-throughput screening of PLGA thin films utilizing hydrophobic fluorescent dyes for hydrophobic drug compounds

2011 ◽  
Vol 100 (10) ◽  
pp. 4317-4329 ◽  
Author(s):  
Terry W.J. Steele ◽  
Charlotte L. Huang ◽  
Saranya Kumar ◽  
Effendi Widjaja ◽  
Freddy Yin Chiang Boey ◽  
...  
Keyword(s):  
Thin Films ◽  
High Throughput ◽  
High Throughput Screening ◽  
Fluorescent Dyes ◽  
Hydrophobic Drug ◽  
Drug Compounds

scholarly journals Involvement of multiple influx and efflux transporters in the accumulation of cationic fluorescent dyes byEscherichia coli

2019 ◽  
Author(s):  
Srijan Jindal ◽  
Lei Yang ◽  
Philip J. Day ◽  
Douglas B. Kell
Keyword(s):  
Steady State ◽  
High Throughput ◽  
High Throughput Screening ◽  
Fluorescent Dyes ◽  
Gene Knockout ◽  
Sybr Green ◽  
Efflux Transporters ◽  
General Strategy ◽  
Individual Gene

AbstractWe used high-throughput flow cytometry to assess the ability of individual gene knockout strains ofE colito take up two membrane-permeable, cationic fluorescent dyes, viz the carbocyanine diS-C3(5) and the DNA dye SYBR Green. Individual strains showed a large range of distributions of uptake. The range of modal steady-state uptakes for the carbocyanine between the different strains was 36-fold. Knockouts of the ATP synthase α- and β-subunits greatly inhibited uptake, implying that most uptake was ATP-driven rather than being driven by say a membrane potential. Dozens of transporters changed the steady-state uptake of the dye by more than 50% with respect to that of the wild type, in both directions (increased or decreased); knockouts in known influx and efflux transporters behaved as expected, giving confidence in the general strategy. Many of the knockouts with the most reduced uptake were transporter genes of unknown function (‘y-genes’). Similarly, several overexpression variants in the ‘ASKA’ collection had the anticipated, opposite effects. Similar findings were made with SYBR Green (the range being some 69-fold), though despite it too containing a benzimidazole motif there was negligible correlation between its uptake and that of the carbocyanine when compared across the various strains. Overall, we conclude that the uptake of these dyes may be catalysed by a great many transporters of possibly broad and presently unknown specificity. This casts serious doubt upon the use of such dyes as quantitative stains for representing either bioenergetic parameters or the amount of cellular DNA in unfixed cells (in vivo). By contrast, it opens up their potential use as transporter assay substrates in high-throughput screening.

Interference: A Much-Neglected Aspect in High-Throughput Screening of Nanoparticles

2020 ◽  
Vol 39 (5) ◽  
pp. 397-421
Author(s):  
Charlene Andraos ◽  
Il Je Yu ◽  
Mary Gulumian
Keyword(s):  
Big Data ◽  
High Throughput ◽  
High Throughput Screening ◽  
Fluorescent Dyes ◽  
Membrane Integrity ◽  
Data Generation ◽  
Label Free ◽  
Predictive Toxicology ◽  
Toxicity Assay ◽  
Novel Concept

Despite several studies addressing nanoparticle (NP) interference with conventional toxicity assay systems, it appears that researchers still rely heavily on these assays, particularly for high-throughput screening (HTS) applications in order to generate “big” data for predictive toxicity approaches. Moreover, researchers often overlook investigating the different types of interference mechanisms as the type is evidently dependent on the type of assay system implemented. The approaches implemented in the literature appear to be not adequate as it often addresses only one type of interference mechanism with the exclusion of others. For example, interference of NPs that have entered cells would require intracellular assessment of their interference with fluorescent dyes, which has so far been neglected. The present study investigated the mechanisms of interference of gold NPs and silver NPs in assay systems implemented in HTS including optical interference as well as adsorption or catalysis. The conventional assays selected cover all optical read-out systems, that is, absorbance (XTT toxicity assay), fluorescence (CytoTox-ONE Homogeneous membrane integrity assay), and luminescence (CellTiter Glo luminescent assay). Furthermore, this study demonstrated NP quenching of fluorescent dyes also used in HTS (2′,7′-dichlorofluorescein, propidium iodide, and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzamidazolocarbocyanin iodide). To conclude, NP interference is, as such, not a novel concept, however, ignoring this aspect in HTS may jeopardize attempts in predictive toxicology. It should be mandatory to report the assessment of all mechanisms of interference within HTS, as well as to confirm results with label-free methodologies to ensure reliable big data generation for predictive toxicology.

scholarly journals Miniaturization of Whole Live Cell-Based GPCR Assays Using Microdispensing and Detection Systems

2004 ◽  
Vol 9 (3) ◽  
pp. 186-195 ◽  
Author(s):  
Oleg Kornienko ◽  
Raul Lacson ◽  
Priya Kunapuli ◽  
Jonathan Schneeweis ◽  
Ira Hoffman ◽  
...  
Keyword(s):  
G Protein ◽  
High Throughput ◽  
High Throughput Screening ◽  
G Protein Coupled Receptors ◽  
Assay Development ◽  
Functional Responses ◽  
Detection Systems ◽  
G Protein Coupled ◽  
Receptor Agonists And Antagonists ◽  
Assay Volume

Cell-based β-lactamase reporter gene assays designed to measure the functional responses of G-protein-coupled receptors (GPCRs) were miniaturized to less than 2 μL total assay volume in a 3456-well microplate. Studies were done to evaluate both receptor agonists and antagonists. The pharmacology of agonists and antagonists for target GPCRs originally developed in a 96-well format was recapitulated in a 3456-well microplate format without compromising data quality or EC50/IC50 precision. These assays were employed in high-throughput screening campaigns, allowing the testing of more than 150,000 compounds in 8 h. The instrumentation used and practical aspects of the assay development are discussed.

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