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High-Throughput Screening for mTORC1/mTORC2 Kinase Inhibitors Using a Chemiluminescence-Based ELISA Assay.
- Source :
- Assay & Drug Development Technologies; Oct2009, Vol. 7 Issue 5, p471-478, 8p
- Publication Year :
- 2009
-
Abstract
- A high-throughput chemiluminescence and ELISA-based biochemical assay to identify mTORC1/mTORC2 kinase inhibitors is described. These mTOR complexes were isolated from HeLa whole cell lysate using mTOR antibodies and in-well immunoprecipitation. The integrity and purity of the mTORC1 and mTORC2 immunocomplexes were confirmed by western blotting. Full-length recombinant 4E-BP1 was used as a substrate and the catalytic activity was measured by detection of p4E-BP1 [T37/46] by a chemiluminescence method. The performance of this assay that can be used to identify dual mTORC1 and mTORC2 kinase inhibitors in a high-throughput 384-well format is described. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 1540658X
- Volume :
- 7
- Issue :
- 5
- Database :
- Complementary Index
- Journal :
- Assay & Drug Development Technologies
- Publication Type :
- Academic Journal
- Accession number :
- 45422680
- Full Text :
- https://doi.org/10.1089/adt.2008.0183