Table 3.
Purification of the 42-kD Kinase
| Fraction | Protein (mg)a |
Total Activity (pmol min −1) |
Specific Activity (pmol min −1 mg−1) |
Recovery (%) |
Purification (fold) |
|---|---|---|---|---|---|
| Total extractb | 3,900 | 135,546 | 34 | 100 | 1 |
| SOURCE 15Q | 38 | 87,150 | 2,305 | 64 | 68 |
| Phenyl-Sepharose | 61 | 49,500 | 8,250 | 36 | 243 |
| Heparin-Sepharose | 0.510 | 34,900 | 68,431 | 26 | 2,013 |
| Mono Q HR 5/5 | 0.025 | 10,150 | 406,000 | 7 | 11,941 |
a
The cell extract was prepared from ∼500 g of BY-2 cells treated for 5 min with 250 mM NaCl.
b
The amount of protein was estimated by comparing different amounts of BSA with the 42-kD protein band on a silver-stained SDS–polyacrylamide gel.