Abstract
A panel of luminescent Saccharomyces cerevisiae cell-based nuclear receptor assays, consisting of human estrogen receptors α and β, androgen receptor, and aryl hydrocarbon receptor, was miniaturized from the standard 96-well microplate format to high throughput 384- and 1536-well microplate formats. In these assays, firefly luciferase lacking the peroxisome targeting sequence was used as a reporter and D-luciferin substrate was pre-mixed with the yeast cells before the incubation step, eliminating cell lysis and substrate addition steps, and allowing multiple readings at any desired time point. All of the assays were highly functional in the 384-well format, and most functioned well in the 1536-well format. The detection limit of the estrogen receptor α assay was even lower in the miniaturized microplate formats than in the original 96-well format. The panel of yeast-cell-based nuclear receptor assays can be used for high throughput chemical testing and environmental monitoring of potential endocrine-disrupting activity of compounds and samples.
Keywords: Cell-based assay, endocrine-disrupting compounds, high throughput assay, luciferase, nuclear receptors, saccharomyces cerevisiae, REACH, high throughput screening, Biomek NXP, Victor3 1420 Multilabel Counter, GraphPad Prism 4, BMAER, BMAAREluc/AR, BMAAhR/ ARNT, estrogen receptor, ER yeast assay, D-luciferin, AR assay, LOD, 17-estradiol, Dihydro- testosterone, Bentzo-a-pyrene, AhR assays, Z' factor
Combinatorial Chemistry & High Throughput Screening
Title: Miniaturization of a Panel of High Throughput Yeast-Cell-Based Nuclear Receptor Assays in 384- and 1536-Well Microplates
Volume: 14 Issue: 1
Author(s): Johanna Rajasarkka and Marko Virta
Affiliation:
Keywords: Cell-based assay, endocrine-disrupting compounds, high throughput assay, luciferase, nuclear receptors, saccharomyces cerevisiae, REACH, high throughput screening, Biomek NXP, Victor3 1420 Multilabel Counter, GraphPad Prism 4, BMAER, BMAAREluc/AR, BMAAhR/ ARNT, estrogen receptor, ER yeast assay, D-luciferin, AR assay, LOD, 17-estradiol, Dihydro- testosterone, Bentzo-a-pyrene, AhR assays, Z' factor
Abstract: A panel of luminescent Saccharomyces cerevisiae cell-based nuclear receptor assays, consisting of human estrogen receptors α and β, androgen receptor, and aryl hydrocarbon receptor, was miniaturized from the standard 96-well microplate format to high throughput 384- and 1536-well microplate formats. In these assays, firefly luciferase lacking the peroxisome targeting sequence was used as a reporter and D-luciferin substrate was pre-mixed with the yeast cells before the incubation step, eliminating cell lysis and substrate addition steps, and allowing multiple readings at any desired time point. All of the assays were highly functional in the 384-well format, and most functioned well in the 1536-well format. The detection limit of the estrogen receptor α assay was even lower in the miniaturized microplate formats than in the original 96-well format. The panel of yeast-cell-based nuclear receptor assays can be used for high throughput chemical testing and environmental monitoring of potential endocrine-disrupting activity of compounds and samples.
Export Options
About this article
Cite this article as:
Rajasarkka Johanna and Virta Marko, Miniaturization of a Panel of High Throughput Yeast-Cell-Based Nuclear Receptor Assays in 384- and 1536-Well Microplates, Combinatorial Chemistry & High Throughput Screening 2011; 14 (1) . https://dx.doi.org/10.2174/1386207311107010047
DOI https://dx.doi.org/10.2174/1386207311107010047 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Computational Characterization of a Series of Eicosanoids
Letters in Drug Design & Discovery Mitochondrial Neuroprotection in Traumatic Brain Injury: Rationale and Therapeutic Strategies
CNS & Neurological Disorders - Drug Targets Brown Recluse Spider Venom: Proteomic Analysis and Proposal of a Putative Mechanism of Action
Protein & Peptide Letters Neuropeptides as Therapeutic Targets to Combat Stress-Associated Behavioral and Neuroendocrinological Effects
CNS & Neurological Disorders - Drug Targets Novel Personalised Drugs for Cystic Fibrosis
Clinical Immunology, Endocrine & Metabolic Drugs (Discontinued) Carbon Nanotube-Based Electrochemical Biosensing Platforms: Fundamentals,Applications, and Future Possibilities
Recent Patents on Biotechnology Recombinant Tissue-Type Plasminogen Activator (Alteplase) in the Management of Acute Ischemic Stroke: A Review of its Pharmacological Properties, Efficacy, Safety and Pharmacoeconomic Aspects
Current Drug Therapy GPR55 and its Interaction with Membrane Lipids: Comparison with Other Endocannabinoid-Binding Receptors
Current Medicinal Chemistry A Review of Pharmacological Treatment Options for Lung Cancer: Emphasis on Novel Nanotherapeutics and Associated Toxicity
Current Drug Targets Editorial [Sentence First, Verdict Afterwards: Using Value of Information Analysis to Inform Decisions about Pharmacogenomic Test Adoption and Research]
Current Pharmacogenomics and Personalized Medicine Transungual Drug Delivery System for the Topical Treatment of Onychomycosis: A Review
Drug Delivery Letters Should We Develop an Inhaled Anti-pneumococcal Vaccine for Adults?
Current Medicinal Chemistry - Anti-Infective Agents Alkaloids and Flavonoids as α1-Adrenergic Receptor Antagonists
Current Medicinal Chemistry Carcinogenesis and Therapeutic Strategies in Thyroid Cancer
Current Drug Targets A Randomized, Placebo-controlled, Triple-blind Study to Determine the Effect of Farlong Ginseng Plus® NotoGinseng Extract on Cholesterol and Blood Pressure
Current Nutraceuticals Engineering of Therapeutic Proteins Production in Escherichia coli
Current Pharmaceutical Biotechnology Postmenopausal Health of Indian Women: A Review
Current Women`s Health Reviews Progress on the Screening and Analysis of Bioactive Compounds in Traditional Chinese Medicines by Biological Fingerprinting Analysis
Combinatorial Chemistry & High Throughput Screening Topics on Pharmacology for Dietitians in Modern Practice
Current Nutrition & Food Science Oxidative Stress in the ICU
Current Nutrition & Food Science