Abstract
The microtubule-associated protein (MAP) tau has been implicated in the pathology of numerous neurodegenerative diseases. In the past decade, the hyperphosphorylated and aggregated states of tau protein have been important targets in the drug discovery field for the potential treatment of Alzheimer’s disease. Although several compounds have been reported to reduce the hyperphosphorylated state of tau or impact the stabilization of tau, their therapeutic activities are remain to be validated. Recently, reduction of total cellular tau protein has emerged as an alternate intervention point for drug development and a potential treatment of tauopathies. We have developed and optimized homogenous assays, using the AlphaLISA and HTRF assay technologies, for the quantification of total cellular tau protein levels in the SH-SY5Y neuroblastoma cell line. The signal-to-basal ratios were 375 and 5.3, and the Z’ factors were 0.67 and 0.60 for the AlphaLISA and HTRF tau assays, respectively. The clear advantages of these homogeneous tau assays over conventional total tau assays, such as ELISA and Western blot, are the elimination of plate wash steps and miniaturization of the assay into 1536-well plate format for the ultra–high-throughput screening of large compound libraries.
Keywords: Alzheimer’ s disease, FRET-based assay, high-throughput screening assay, prions, neurodegenerative diseases, tau protein.
Current Alzheimer Research
Title:A High-Throughput Screening Assay for Determining Cellular Levels of Total Tau Protein
Volume: 10 Issue: 7
Author(s): Seameen J. Dehdashti, Wei Zheng, Joel R. Gever, Robert Wilhelm, Dac-Trung Nguyen, Gurusingham Sittampalam, John C. McKew, Christopher P. Austin and Stanley B. Prusiner
Affiliation:
Keywords: Alzheimer’ s disease, FRET-based assay, high-throughput screening assay, prions, neurodegenerative diseases, tau protein.
Abstract: The microtubule-associated protein (MAP) tau has been implicated in the pathology of numerous neurodegenerative diseases. In the past decade, the hyperphosphorylated and aggregated states of tau protein have been important targets in the drug discovery field for the potential treatment of Alzheimer’s disease. Although several compounds have been reported to reduce the hyperphosphorylated state of tau or impact the stabilization of tau, their therapeutic activities are remain to be validated. Recently, reduction of total cellular tau protein has emerged as an alternate intervention point for drug development and a potential treatment of tauopathies. We have developed and optimized homogenous assays, using the AlphaLISA and HTRF assay technologies, for the quantification of total cellular tau protein levels in the SH-SY5Y neuroblastoma cell line. The signal-to-basal ratios were 375 and 5.3, and the Z’ factors were 0.67 and 0.60 for the AlphaLISA and HTRF tau assays, respectively. The clear advantages of these homogeneous tau assays over conventional total tau assays, such as ELISA and Western blot, are the elimination of plate wash steps and miniaturization of the assay into 1536-well plate format for the ultra–high-throughput screening of large compound libraries.
Export Options
About this article
Cite this article as:
Dehdashti J. Seameen, Zheng Wei, Gever R. Joel, Wilhelm Robert, Nguyen Dac-Trung, Sittampalam Gurusingham, McKew C. John, Austin P. Christopher and Prusiner B. Stanley, A High-Throughput Screening Assay for Determining Cellular Levels of Total Tau Protein, Current Alzheimer Research 2013; 10 (7) . https://dx.doi.org/10.2174/15672050113109990143
DOI https://dx.doi.org/10.2174/15672050113109990143 |
Print ISSN 1567-2050 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5828 |
Call for Papers in Thematic Issues
New Advances in the Prevention, Diagnosis, Treatment, and Rehabilitation of Alzheimer's Disease
Aims and Scope: Introduction: Alzheimer's disease (AD) poses a significant global health challenge, with an increasing prevalence that demands concerted efforts to advance our understanding and strategies for prevention, diagnosis, treatment, and rehabilitation. This thematic issue aims to bring together cutting-edge research and innovative approaches from multidisciplinary perspectives to address ...read more
Current updates on the Role of Neuroinflammation in Neurodegenerative Disorders
Neuroinflammation is an invariable hallmark of chronic and acute neurodegenerative disorders and has long been considered a potential drug target for Alzheimer?s disease (AD) and dementia. Significant evidence of inflammatory processes as a feature of AD is provided by the presence of inflammatory markers in plasma, CSF and postmortem brain ...read more
Deep Learning for Advancing Alzheimer's Disease Research
Alzheimer's disease (AD) poses a significant global health challenge, with an increasing number of individuals affected yearly. Deep learning, a subfield of artificial intelligence, has shown immense potential in various domains, including healthcare. This thematic issue of Current Alzheimer Research explores the application of deep learning techniques in advancing our ...read more
Diagnostic and therapeutic biomarkers of dementia
Dementia affects 18 million people worldwide. Dementia is a syndrome of symptoms caused by brain disease, usually chronic or progressive, clinically characterized by multiple impairments of higher cortical functions such as memory, thinking, orientation, and learning. In addition, in the course of dementia, cognitive deficits are observed, which often hinder ...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
- Announcements
Related Articles
-
ChemoImmunoModulation: Immune Regulation by the Antineoplastic Chemotherapeutic Agents
Current Medicinal Chemistry The Therapeutic Target Hsp90 and Cancer Hallmarks
Current Pharmaceutical Design Recent Advances in Minimal Access Surgery for Infants and Children
Current Pediatric Reviews Silencing of Disease-related Genes by Small Interfering RNAs
Current Molecular Medicine The ARRONAX Project
Current Radiopharmaceuticals Natural Products as Promising Drug Candidates for the Treatment of Alzheimer’s Disease: Molecular Mechanism Aspect
Current Neuropharmacology Synthesis and Biological Activity of Unnatural Enediynes
Current Medicinal Chemistry Identification of Fish Cell Lines Using 2-D Electrophoresis Based Protein Expression Signatures
Current Proteomics A Medicinal Chemist’s Perspective Towards Structure Activity Relationship of Heterocycle Based Anticancer Agents
Current Topics in Medicinal Chemistry Breast Cancer: A Review of Risk Factors and New Insights into Treatment
Current Cancer Therapy Reviews Plant Defensins and Defensin-Like Peptides - Biological Activities and Biotechnological Applications
Current Pharmaceutical Design Tankyrase as a Novel Molecular Target in Cancer and Fibrotic Diseases
Current Drug Targets Therapeutical Potential of CB<sub>2</sub> Receptors in Immune-Related Diseases
Current Molecular Pharmacology Hormonal Control of the Neuropeptide Y System
Current Protein & Peptide Science Chemotherapy In Japanese Encephalitis: Are We There Yet?
Infectious Disorders - Drug Targets Promising Anti-Fibrotic Approaches for Future Treatment of Systemic Sclerosis
Current Rheumatology Reviews NRF2-Dependent Glutamate-L-Cysteine Ligase Catalytic Subunit Expression Mediates Insulin Protection Against Hyperglycemia-Induced Brain Endothelial Cell Apoptosis
Current Neurovascular Research The Roles of Histone Demethylase UTX and JMJD3 (KDM6B) in Cancers: Current Progress and Future Perspectives
Current Medicinal Chemistry The Involvement of the Vasopressin System in Stress-Related Disorders
CNS & Neurological Disorders - Drug Targets Breast Cancer Stem Cells and Intrinsic Subtypes: Controversies Rage On
Current Stem Cell Research & Therapy