| Large-scale analysis of UPR-mediated apoptosis in human cells. | |
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MedLine Citation:
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PMID: 21329794 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The historic distinction between academic- and industry-driven drug discovery, whereby academicians worked to identify therapeutic targets and pharmaceutical companies advanced probe discovery, has been blurred by an academic high-throughput chemical genomic revolution. It is now common for academic labs to use biochemical or cell-based high-throughput screening (HTS) to investigate the effects of thousands or even hundreds of thousands of chemical probes on one or more targets over a period of days or weeks. To support the efforts of individual investigators, many universities have established core facilities where screening can be performed collaboratively with large chemical libraries managed by highly trained HTS personnel and guided by the experience of computational, medicinal, and synthetic organic chemists. The identification of large numbers of promising hits from such screens has driven the need for independent labs to scale down secondary in vitro assays in the hit to lead identification process. In this chapter, we will describe the use of luminescent and quantitative reverse transcription real-time PCR (qRT-PCR) technologies that permit evaluation of the expression patterns of multiple unfolded protein response (UPR) and apoptosis-related genes, and simultaneously evaluate proliferation and cell death in 96- or 384-well format. |
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Authors:
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Andrew M Fribley; Justin R Miller; Tyler E Reist; Michael U Callaghan; Randal J Kaufman |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Methods in enzymology Volume: 491 ISSN: 1557-7988 ISO Abbreviation: Meth. Enzymol. Publication Date: 2011 |
Date Detail:
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Created Date: 2011-02-18 Completed Date: 2011-05-24 Revised Date: 2012-05-15 |
Medline Journal Info:
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Nlm Unique ID: 0212271 Medline TA: Methods Enzymol Country: United States |
Other Details:
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Languages: eng Pagination: 57-71 Citation Subset: IM |
Copyright Information:
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Copyright © 2011 Elsevier Inc. All rights reserved. |
Affiliation:
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Department of Biological Chemistry, University of Michigan School of Medicine, Ann Arbor, Michigan, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Apoptosis* Cell Line Cytological Techniques / methods DNA Fragmentation Gene Expression Profiling / methods* Humans Polymerase Chain Reaction / methods Unfolded Protein Response* |
| Grant Support | |
ID/Acronym/Agency:
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DE019678/DE/NIDCR NIH HHS; DK042394/DK/NIDDK NIH HHS; HL052173/HL/NHLBI NIH HHS; HL057346/HL/NHLBI NIH HHS; MH084182/MH/NIMH NIH HHS; MH089782/MH/NIMH NIH HHS; R01 DK088227-02/DK/NIDDK NIH HHS; R01 HL052173/HL/NHLBI NIH HHS; R01 HL052173-14A1/HL/NHLBI NIH HHS; R03 MH089782-02/MH/NIMH NIH HHS; R37 DK042394-14/DK/NIDDK NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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