Table 6.
Summary of Sts-1 HTS assay protocol for 1,536-well plate format
| Step | Description | Quantity/Condition | Comments |
|---|---|---|---|
| 1 | Sts-1HP addition in buffer | 4 μL/well | 750 ng/mL STS-1 in Assay buffer (30 mM Tris HCl, 75 mM NaCl, 1 mM EDTA, 1 mM DTT) |
| 2 | Pin compound or DMSO | 35 nL/well | 5.8 μM compound in 0.4% DMSO final |
| 3 | OMFP addition in water | 2 μL/well | 50uM final |
| 4 | Incubation | 20 minutes | Room temperature in the dark |
| 5 | NaOH addition | 1 μL/well | 100 mM final, stops reaction |
| 6 | Read fluorescence | Viewlux | Ex: 480 nm; Em: 520 nm |