Table 1.
Step of purification | Protein (mg) | Total activity (pmol min−1) | Specific activity (pmol min−1 mg−1) | Fold of purification | Recovery (%) |
Homogenatea | 3740 | 7106 | 2 | 1 | 100.0 |
30% (NH4)2SO4 | 258 | 4489 | 17 | 9 | 63.2 |
100 000 g | 171 | 3454 | 20 | 10 | 48.6 |
Q-Sepharose FF | 11.56 | 983 | 85 | 43 | 13.8 |
Phenyl-Sepharose FF (HS) | 1.73 | 415 | 240 | 120 | 5.8 |
Resource Q | 0.368 | 307 | 834 | 417 | 4.3 |
Mono Q™ 5/50 GLb | 0.138 | 249 | 1 804 | 902 | 3.5 |
Poly-L-lysine-agaroseb | 0.021 | 189 | 9 005 | 4 502 | 2.7 |
Hiload 16/60Superdex 75pgb | 0.004 | 82 | 20 420 | 10 210 | 1.1 |
The starting homogenate was prepared from 1000 g of maize leaves treated with 100 μM ABA for 2 h. MBP was used as substrate in the kinase assays. Total protein amount was measured according to Bradford (1976).
The amount of protein was estimated by comparison with known bovine serum albumin on a Coomassie-stained SDS–polyacrylamide gel.