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Assay Method Information

Assay Name:  Dose response counterscreen of uHTS hits for ATG4B inhibitors in a Phospholipase A2 assay
Description:  Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH090871-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA This assay is a counterscreen for the primary screen (AID504462) which looked for inhibitors of Autophagin 1 (ATG4B). In screening for compounds that inhibit ATG4B, the High Throughput Screening (HTS) assay utilized a cleavable form of Phospholipase A2 (PLA2), which is expressed as a fusion protein with the Autophagin substrate LC3/ATG8 appended to its N-terminus, as the substrate for the primary enzymatic reaction. The addition of sequences to the N-terminus of PLA2 inhibits the activity of this enzyme. Cleavage by proteases removing the N-terminal extension then restores enzyme activity, constituting the basis for a protease assay. The sub
Affinity data for this assay
 

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Last update November 1, 2007
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