Purification Scheme for Cofactor B
| Dimension . | . | Matrix . | . | Size*:Buffer‡ . | . | Activity‖ . |
|---|---|---|---|---|---|---|
| 1 | Q-Sepharose HP | 5/25:I/II | 80–100 mM | |||
| 2 | Hydroxylapatite | 2.6/12:III/IV | nonbinding | |||
| 3 | MonoQ | 1/10:XIV/XV | 0 .22–0.24 M | |||
| 4 | Q-Sepharose HP | 1/10:XIX/XX§ | 0 .25–0.26 M | |||
| 5 | TSKG3000SW | 0.75/60:X |
| Dimension . | . | Matrix . | . | Size*:Buffer‡ . | . | Activity‖ . |
|---|---|---|---|---|---|---|
| 1 | Q-Sepharose HP | 5/25:I/II | 80–100 mM | |||
| 2 | Hydroxylapatite | 2.6/12:III/IV | nonbinding | |||
| 3 | MonoQ | 1/10:XIV/XV | 0 .22–0.24 M | |||
| 4 | Q-Sepharose HP | 1/10:XIX/XX§ | 0 .25–0.26 M | |||
| 5 | TSKG3000SW | 0.75/60:X |
Size of column bed (width/height) in cm.
Buffer composition as described in Tian et al. (1996)
The composition of buffers used in this dimension was XIX: 20 mM diethanolamine-HCl, 20 mM KCl, 1 mM MgCl2, 1 mM EGTA, 1 mM DTT; XX: same as XIX, except containing 1 M in place of 20 mM KCl.
Counterion concentration containing cofactor B activity.