Table

Table 1. Buffer systems for broad and narrow pH intervals in chromatofocusing with Mono P columns

Approximate volumes (mL)
Mono P 5/20 Mono P 5/5
pH range Start buffer (mL) Eluent (100 mL) Total eluent Pregradient Total eluent Pregradient
Buffer systems for broad pH intervals
9-7 25 mM diethanolamine (pH 9.5), HCl 1.0 mL Pharmalyte (pH 8-10.5), 5.2 mL Polybuffer 96 (pH 7), HCl 34 7 11 2
9-6 25 mM diethanolamine (pH 9.5), HCl 10 mL Polybuffer 96 (pH 6), HCl 34 9 19 2
9-6 25 mM Tris (pH 9.3), acetic acid 10 mL Polybuffer 96 (pH 6), acetic acid 30 3 17 2
8-6 25 mM triethanolamine (pH 8.3), acetic acid 0.21 mL Pharmalyte (pH 8-10.5), 9 mL Polybuffer 96 (pH 6), acetic acid 37 7 15 4
8-5 25 mM triethanolamine (pH 8.3), iminodiacetic acida 3 mL Polybuffer 96, 7 mL Polybuffer 74 (pH 5), iminodiacetic acida 47 6 15 3
7-5 25 mM Bis-Tris (pH 7.1), HCl 10 mL Polybuffer 74 (pH 5), HCl 26 3 13 3
7-4 25 mM Bis-Tris (pH 7.1), iminodiacetic acida 10 mL Polybuffer 74 (pH 4), iminodiacetic acida 46 3 19 3
6-4 25 mM Bis-Tris (pH 6.3), HCl 10 mL Polybuffer 74 (pH 4), HCl 39 3 16 3
Buffer systems for narrow pH intervals
9-8 25 mM diethanolamine (pH 9.4), HCl 1.0 mL Pharmalyte (pH 8-10.5), 5.2 mL Polybuffer 96 (pH 8), HCl 28 3 10 3
8.5-7.5 25 mM Tris (pH 8.8), acetic acid 0.11 mL Pharmalyte (pH 8-10.5), 9.5 mL Polybuffer 96 (pH 7.5), acetic acid 29 4 10 4
8-7 25 mM triethanolamine (pH 8.3), HCl 10 mL Polybuffer 96 (pH 7), HCl 29 5 10 4
7.5-6.5 25 mM methylimidazole (pH 7.6), acetic acid 10 mL Polybuffer 96 (pH 6.5), acetic acid 27 9 11 6
7-6 25 mM Bis-Tris (pH 7), acetic acid 9.5 mL Polybuffer 96, 0.5 mL Polybuffer 74 (pH 6), acetic acid 28 10 12 5
6.5-5.5 25 mM Bis-Tris (pH 6.7), acetic acid 4 mL Polybuffer 96, 6 mL Polybuffer 74 (pH 5.5), acetic acid 23 5 9 3
6-5 25 mM Bis-Tris (pH 6.7), HCl 10 mL Polybuffer 74 (pH 5), HCl 25 3 10 3
5.5-4.5 25 mM piperazine (pH 6.3), HCl or iminodiacetic acida 10 mL Polybuffer 74 (pH 4.5), HCl or iminodiacetic acida 24 3 10 3
5-4 25 mM methylpiperazine (pH 5.7) or iminodiacetic acida 10 mL Polybuffer 74 (pH 4), HCl or iminodiacetic acida 27 7 11 3
For details on buffer preparation, see Purification of Proteins by Chromatofocusing (Mohammad 2010c).
aUse a saturated solution of iminodiacetic acid.
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  1. doi:10.1101/pdb.tab195376 Cold Spring Harb Protoc 2010: pdb.tab195376-
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