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Updated: Jul 18, 2026
12:11
A Rhodopsin Transport Assay by High-Content Imaging Analysis
Published on: January 16, 2019
Protein translocation assays: key tools for accessing new biological information with high-throughput microscopy
Arne Heydorn1, Betina K Lundholt, Morten Praestegaard
1Fisher BioImage ApS, Søborg, Denmark.
Methods in Enzymology
|November 18, 2006
Summary
Redistribution technology offers high-content, high-throughput cell-based assays for drug discovery. Novel applications include a p53:hdm2 assay for hit selection and siRNA screening for pathway analysis.
Area of Science:
- Cellular biology
- Assay development
- Drug discovery
Background:
- Redistribution technology utilizes protein translocation to monitor cellular signaling pathways.
- This high-content, high-throughput assay is valuable for lead profiling and screening.
- Commercially available instruments facilitate its application.
Purpose of the Study:
- To describe two novel high-content Redistribution assay technologies.
- To demonstrate the utility of these assays in drug discovery and pathway analysis.
Main Methods:
- Protein targets are labeled with green fluorescent protein in stably transfected cell lines.
- Assays are read using high-throughput optical microscopy.
- Two novel assays are presented: p53:hdm2 GRIP interaction assay and siRNA application in FKHR translocation assay.
Main Results:
- The p53:hdm2 assay uses distinct image features for hit detection and mode-of-action deselection.
- siRNA-mediated knockdown of Akt isoforms in a FKHR translocation assay effectively reports on PI3 kinase signaling.
Conclusions:
- Redistribution assays provide powerful tools for high-content screening and pathway analysis.
- These novel applications enhance the utility of Redistribution technology in drug discovery.
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