Table 2.
Purification of human P450 4F11 from E. coli membrane fractions.
| Purification step | P450 (nmol) | Yield (%) |
|---|---|---|
| Membranes | 227 | 100 |
| Solubilized fractiona | 214 | 94 |
| Ni2+-NTA column fractionb | 106 | 47 |
| Hydroxylapatite column fractionc | 73 | 32 |
Solubilization using a mixture of sodium cholate plus Tergitol NP-10, followed by centrifugation to remove insoluble material.
P450 eluted with 200 mM imidazole.
Eluted with 0.7 M potassium phosphate buffer (pH 7.7) containing 0.2% sodium cholate (w/v).