ArticleAssay and drug development technologies2016
Diaphorase Coupling Protocols for Red-Shifting Dehydrogenase Assays.
Article in Assay and drug development technologies, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 22 citations in OpenAlex.
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- Biocatalytic- and Chemoproteomic-Guided Discovery of a PHGDH Inhibitor from Chemoenzymatic-Promoted DNA-Encoded Libraries Selection Platform.Journal of the American Chemical Society · 2025Article
- Integrated Approach of Machine Learning and High-Throughput Screening to Identify Chemical Probe Candidates Targeting Aldehyde Dehydrogenases.ACS pharmacology & translational science · 2025Article
- Optimization of a high throughput screening platform to identify inhibitors of asymmetric diadenosine polyphosphatases.Analytical biochemistry · 2025Article
- Discovery and Characterization of a Novel Allosteric Small-Molecule Inhibitor of NADPBiochemistry · 2022Article
- Fucosidases from the human gut symbiont Ruminococcus gnavus.Cellular and molecular life sciences : CMLS · 2021Article
- Identification of Activators of Human Fumarate Hydratase by Quantitative High-Throughput Screening.SLAS discovery : advancing life sciences R & D · 2020Article
- Paper-Based Resazurin Assay of Inhibitor-Treated Porcine Sperm.Micromachines · 2019Article
- Discovery and optimization of piperazine-1-thiourea-based human phosphoglycerate dehydrogenase inhibitors.Bioorganic & medicinal chemistry · 2018Article
- Discovery and Optimization of Potent, Cell-Active Pyrazole-Based Inhibitors of Lactate Dehydrogenase (LDH).Journal of medicinal chemistry · 2017Article
- A new non-radioactive deoxyhypusine synthase assay adaptable to high throughput screening.Amino acids · 2017Article
- Assessing inhibitors of mutant isocitrate dehydrogenase using a suite of pre-clinical discovery assays.Scientific reports · 2017Article
- Small-Molecule Screens: A Gateway to Cancer Therapeutic Agents with Case Studies of Food and Drug Administration-Approved Drugs.Pharmacological reviews · 2017Review
- A High-Content Assay Enables the Automated Screening and Identification of Small Molecules with Specific ALDH1A1-Inhibitory Activity.PloS one · 2017Article
- Avoiding Fluorescence Assay Interference-The Case for Diaphorase.Assay and drug development technologies · 2016Review
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Dehydrogenases are an important target for the development of cancer therapeutics. Dehydrogenases either produce or consume NAD(P)H, which is fluorescent but at a wavelength where many compounds found in chemical libraries are also fluorescent. By coupling dehydrogenases to diaphorase, which utilizes NAD(P)H to produce the fluorescent molecule resorufin from resazurin, the assay can be red-shifted into a spectral region that reduces interference from compound libraries. Dehydrogenases that produce NAD(P)H, such as isocitrate dehydrogenase 1 (IDH1), can be read in kinetic mode. Dehydrogenases that consume NAD(P)H, such as mutant IDH1 R132H, can be read in endpoint mode. Here, we report protocols for robust and miniaturized 1,536-well assays for WT IDH1 and IDH1 R132H coupled to diaphorase, and the counterassays used to further detect compound interference with the coupling reagents. This coupling technique is applicable to dehydrogenases that either produce or consume NAD(P)H, and the examples provided here can act as guidelines for the development of high-throughput screens against this enzyme class.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.