| Solution Information | help | |
| Enzyme: | Plectin | |
| inhibitor: | BDBM54771 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) Rhodamine-Plectin and Siah provided by the assay provider 2) Assay buffer: 25mM Bis-Tris pH 7.0, 1mM TCEP and 0.005% Tween 20 3) Siah/Rd-Plectin Assay mix: 0.005uM Rd-Plectin, 0.016uM SIAH in Assay Buffer uHTS Protocol 1) Add 6 uL of Rd-Plectin (Positive Control) to Columns 1-2 of a black, opaque, Corning 1536-well plate (Cat#3724) using a Multidrop Combi. 2) Add 6 uL of Siah/Rd Plectin Assay Mix to Columns 3-48 using a Multidrop Combi. 3) Using a HighResolution Biosolutions pintool dispense 60nl of: 100% DMSO to columns 1-4 (1% final) 2mM Compounds in 100% DMSO to columns 5-48 (20uM final) 4) Following a 1 hr incubation at room temperature, in the dark, the plates are read on a BMG Pherastar using Rhodamine FP filters. Dose Response Protocol 1) Add 6 uL of Rd-Plectin (Positive Control) to Columns 1-2 of a black, opaque, Corning 1536-well plate (Cat#3724) using a Multidrop Combi. 2) Add 6 uL of Siah/Rd Plectin Assay Mix to Columns 3-48 using a Multidrop Combi. 3) | |
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