| Solution Information | help | |
| Enzyme: | Cysteine protease ATG4B | |
| inhibitor: | BDBM42035 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | A. Brief Description of the Assay: This assay measures the activity of inhibitors of ATG4B enzyme that are able to prevent cleavage of LC3-PLA2. It will be measured by Fluorescence in 1536 well plate format. B. Materials: Item, Source, Cat# ATG4B Enzyme, Assay Provider LC3-PLA2, Assay Provider NBD-C6-HPC, Sigma-Aldrich, N3786) TRIS-HCl pH 8.0 CaCl2, Sigma-Aldrich, 499609 Dithiothreitol, Sigma-Aldrich, 646563 1536-well Black solid bottom (Normal-Binding) plate, Greiner, 782076 C. Procedures: 1. Using LabCyte Echo, transfer appropriate volumes of test compounds in DMSO from an Echo qualified source plate to an assay plate (Col. 5 - 48) in order to achieve appropriate test concentrations and range. Backfill with DMSO to equilibrate DMSO concentration across test compound wells. Add equal volume of DMSO to control wells (Col. 1-4). 2. Prepare reagents as stated in D. Recipe and keep on ice. Note: Reagents should be used as soon as possible. Final signal will degrade by half over 1.5 h | |
| If you find an error in this entry please send us an E-mail | ||