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Preparation of FRET reporters to support chemical probe development.

ABSTRACT: In high throughput screening (HTS) campaigns, the quality and cost of commercial reagents suitable for pilot studies often create obstacles upon scale-up to a full screen. We faced such challenges in our efforts to implement an HTS for inhibitors of the phosphopantetheinyl transferase Sfp using an assay that had been validated using commercially available reagents. Here we demonstrate a facile route to the synthetic preparation of reactive tetraethylrhodamine and quencher probes, and their application to economically produce fluorescent and quencher-modified substrates. These probes were prepared on a scale that would allow a full, quantitative HTS of more than 350,000 compounds.

SUBMITTER: Foley TL

PROVIDER: S-EPMC2948233 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature

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Preparation of FRET reporters to support chemical probe development.

Foley Timothy L TL Yasgar Adam A Garcia Christopher J CJ Jadhav Ajit A Simeonov Anton A Burkart Michael D MD

Organic & biomolecular chemistry 20100820 20

In high throughput screening (HTS) campaigns, the quality and cost of commercial reagents suitable for pilot studies often create obstacles upon scale-up to a full screen. We faced such challenges in our efforts to implement an HTS for inhibitors of the phosphopantetheinyl transferase Sfp using an assay that had been validated using commercially available reagents. Here we demonstrate a facile route to the synthetic preparation of reactive tetraethylrhodamine and quencher probes, and their appli ...[more]

PMID: 20725690

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