| Solution Information | help | |
| Enzyme: | Plectin | |
| inhibitor: | BDBM54902 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) Fluorescein-Plectin and Siah provided by the assay provider 2) Terbium His Antibody was obtained from Invitrogen 3) Assay buffer: 25mM Bis-Tris pH 7.0, 1mM TCEP and 0.005% Tween 20 4) Siah - TB His Antibody Mix: 10 nM Siah, 5 nM Tb His Antibody in Assay Buffer 5) Positive Control: 2.5 nM Tb His Antibody, 5 nM Fl-Plectin in Assay Buffer 6) Fl-Plectin Solution: 10 nM Fl-Plectin Dose Response Protocol 1) Add 6 uL of Positive Control to Columns 1-2 of a black, opaque, Corning 1536-well plate (Cat#3724) using a Multidrop Combi. 2) Combine in equal parts the Siah - Tb His Antibody Mix with the Fl-Plectin solution. 3) Add 6 uL of Siah - Tb His Antibody/Fl-Plectin solution Mix to Columns 3-48 using a Multidrop Combi. 4) Using a Labcyte Echo, DMSO and test compounds are transferred to wells. DMSO only is transferred to columns 1-3 and 46-48(Control wells), while varying volumes of test compounds are transferred to columns 4-45 to achieve the desired test concentrations. | |
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