Binding Buffer for Mobility Shift Assays
| Reagent | Quantity (for 10 mL) | Final concentration |
|---|---|---|
| Tris (1 m; pH 8.0) | 0.4 mL | 40 mm |
| KCl (2 m) | 0.15 mL | 30 mm |
| MgCl2 (1 m) | 10 µL | 1 mm |
| NP40 (10%, w/v) | 10 µL | 0.01% (w/v) |
| Dithiothreitol (DTT) (1 m) | 10 µL | 1 mm |
| H2O or extract | 9.42 mL | |
| If the proteins or extract used for binding are not in glycerol, add 5% to the binding buffer. Store indefinitely at −20°C. | ||
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