Table 1. General schemes for eluting proteins from hydroxyapatite
| Step elution | Gradient elution |
|---|---|
| Scheme I | |
| 1. 0.005 M MgCl2 to elute basic proteins | 0.001 to 0.005 M MgCl2 gradient |
| 2. 1.0 M MgCl2 to elute proteins with pI values between 5 and 8 | 1.0 M MgCl2 in a step |
| 3. 0.3 M PO4 to elute acidic proteins | 0.01 to 0.3 M PO4 gradient |
| Scheme II | |
| 1. 0.005 M MgCl2 to elute basic proteins | 0.001 to 0.005 M MgCl2 gradient |
| 2. 1 M NaCl or KCl to elute proteins with pI values of 7.0 to 7.6 and DNases | 0.01 to 1.0 M NaCl or KCl gradient |
| 3. 0.3 M PO4 to elute acidic proteins | 0.01 to 0.3 M PO4 gradient |
| Scheme III | |
| 1. 1 M NaCl or KCl to elute basic and neutral proteins, as well as DNases | 1 M NaCl or KCl in a step |
| 2. 0.3 M PO4 to elute acidic proteins | 0.01 to 0.3 M PO4 gradient |
Reproduced from Grobunoff (1990) with permission.
In the three elution schemes outlined in this table, proteins will emerge in the following order from a column: basic, neutral, acidic
Table 1.
General schemes for eluting proteins from hydroxyapatite
