Table 1.1.
Unfolding reaction setup
| Urea (μL) |
Phosphate buffer (μL) |
DTT (μL) |
Native protein (μL) |
Final urea [M] |
|---|---|---|---|---|
| 0 | 985 | 10 | 5 | 0 |
| 50 | 935 | 10 | 5 | 0.5 |
| 100 | 885 | 10 | 5 | 1.0 |
| 150 | 835 | 10 | 5 | 1.5 |
| 200 | 785 | 10 | 5 | 2.0 |
| 250 | 735 | 10 | 5 | 2.5 |
| 300 | 685 | 10 | 5 | 3.0 |
| 350 | 635 | 10 | 5 | 3.5 |
| 400 | 585 | 10 | 5 | 4.0 |
| 450 | 535 | 10 | 5 | 4.5 |
| 500 | 485 | 10 | 5 | 5.0 |
| 550 | 435 | 10 | 5 | 5.5 |
| 600 | 385 | 10 | 5 | 6.0 |
| 650 | 335 | 10 | 5 | 6.5 |
| 700 | 285 | 10 | 5 | 7.0 |
| 750 | 235 | 10 | 5 | 7.5 |
| 800 | 185 | 10 | 5 | 8.0 |
Calculations are based on stock concentrations of 200 μM native protein, 10 M urea, 100 mM DTT, and 50 mM phosphate buffer, pH 7.5. The final protein concentration is 1 μM.