| 10.5-9a |
-- |
-- |
-- |
-- |
-- |
-- |
-- |
| 10.5-8 |
PBE 118 |
pH 11, 25 mM triethylamine-HCl |
pH 8 Pharmalyte (pH 8-10.5)-HCl |
1:45 |
1.5 |
11.5 |
13.0 |
| 10.5-7 |
PBE 118 |
pH 11, 25 mM triethylamine-HCl |
pH 7 Pharmalyte (pH 8-10.5)-HCl |
1:45 |
2.0 |
11.5 |
13.5 |
| 9-8b |
PBE 94 |
pH 9.4, 25 mM ethanolamine-HCl |
pH 8 Pharmalyte (pH 8-10.5)-HCl |
1:45 |
1.5 |
10.5 |
12.0 |
| 9-7 |
PBE 94 |
pH 9.4, 25 mM ethanolamine-HCl |
pH 7 Polybuffer 96-HCl |
1:10 |
2.0 |
12.0 |
14.0 |
| 9-6 |
PBE 94 |
pH 9.4, 25 mM ethanolamine-acetic acid |
pH 6 Polybuffer 96-acetic acid |
1:10 |
1.5 |
10.5 |
12.0 |
| 8-7 |
PBE 94 |
pH 8.3, 25 mM Tris-HCl |
pH 7 Polybuffer 96-HCl |
1:13 |
1.5 |
9.0 |
10.5 |
| 8-6 |
PBE 94 |
pH 8.3, 25 mM Tris-acetic acid |
pH 6 Polybuffer 96-acetic acid |
1:13 |
3.0 |
9.0 |
12.0 |
| 8-5c |
PBE 94 |
pH 8.3, 25 mM Tris-acetic acid |
pH 5 Polybuffer 96 (30%) plus Polybuffer 74 (70%)-acetic acid |
1:10 |
2.0 |
8.5 |
10.5 |
| 7-6 |
PBE 94 |
pH 7.4, 25 mM imidazole acetic acid |
pH 6 Polybuffer 96-acetic acid |
1:13 |
3.0 |
7.0 |
10.0 |
| 7-5 |
PBE 94 |
pH 7.4, 25 mM imidazole-HCl |
pH 5 Polybuffer 74-HCl |
1:8 |
2.5 |
11.5 |
14.0 |
| 7-4 |
PBE 94 |
pH 7.4, 25 mM imidazole-HCl |
pH 4 Polybuffer 74-HCl |
1:8 |
2.5 |
11.5 |
14.0 |
| 6-5 |
PBE 94 |
pH 6.2, 25 mM histidine-HCl |
pH 5 Polybuffer 74-HCl |
1:10 |
2.0 |
8.0 |
10.0 |
| 6-4 |
PBE 94 |
pH 6.2, 25 mM histidine-HCl |
pH 4 Polybuffer 74-HCl |
1:8 |
2.0 |
7.0 |
9.0 |
| 5-4 |
PBE 94 |
pH 5.5, 25 mM piperazine-HCl |
pH 4 Polybuffer 74-HCl |
1:10 |
3.0 |
9.0 |
12.0 |
|
| For details on buffer preparation, see Purification of Proteins by Chromatofocusing (Mohammad 2010d).
|
| aGradients ending at pH 9 are not recommended, because 9 is above the pH of Pharmalyte (pH 8-10.5).
|
| bPBE 118 and Pharmalyte pH 8-10.5 also cover this range, as do PBE 94 and Polybuffer 96.
|
| cMixing gives the best results. Polybuffer 74 works better than Polybuffer 96 when used alone.
|
