Separation Options with Sephacryl®

Extracted from Size Exclusion Chromatography Principles and Methods, (PDF)
GE Healthcare, 2014

Five Sephacryl® HR chromatography media are available as prepacked columns and in bulk packs (Table 4.1).

Table 4.1. Separation options with Sephacryl® media

Product Fractionation range, Mr (globular proteins) Fractionation range, Mr (dextrans) Sample loading capacity* Maximum pressure drop over the packed bed Recommended operation flow†
HiPrep™ 16/60 Sephacryl® S-100 HR 1 × 103 to 1 × 105   ≤ 5 ml 0.15 MPa, 1.5 bar,21 psi 0.5 ml/min
HiPrep™ 26/60Sephacryl® S-100 HR 1 × 103 to 1 × 105   ≤ 13 ml 0.15 MPa, 1.5 bar,21 psi 1.3 ml/min
Sephacryl® S-100 HR(Bulk medium) 1 × 103 to 1 × 105   0.5% to 4% of total column volume Column-dependent 10 to 35 cm/h
HiPrep™ 16/60Sephacryl® S-200 HR 5 × 103 to 2.5 × 105 1 × 103 to 8 × 104 ≤ 5 ml 0.15 MPa, 1.5 bar,21 psi 0.5 ml/min
HiPrep™ 26/60Sephacryl® S-200 HR 5 × 103 to 2.5 × 105 1 × 103 to 8 × 104 ≤ 13 ml 0.15 MPa, 1.5 bar,21 psi 1.3 ml/min
Sephacryl® S-200 HR (Bulk medium) 5 × 103 to 2.5 × 105 1 × 103 to 8 × 104 0.5% to 4% of total column volume Column-dependent 10 to 35 cm/h
HiPrep™ 16/60Sephacryl® S-300 HR 1 × 104 to 1.5 × 106 2 × 103 to 4 × 105 ≤ 5 ml 0.15 MPa, 1.5 bar,21 psi 0.5 ml/min
HiPrep™ 26/60Sephacryl® S-300 HR 1 × 104 to 1.5 × 106 2 × 103 to 4 × 105 ≤ 13 ml 0.15 MPa, 1.5 bar,21 psi 1.3 ml/min
Sephacryl® S-300 HR(Bulk medium) 1 × 104 to 1.5 × 106 2 × 103 to 4 × 105 0.5% to 4% of total column volume Column-dependent 10 to 35 cm/h
HiPrep™ 16/60Sephacryl® S-400 HR 2 × 104 to 8 × 106 1 × 104 to 2 × 106 ≤ 5 ml 0.15 MPa, 1.5 bar,21 psi 0.5 ml/min
HiPrep™ 26/60Sephacryl® S-400 HR 2 × 104 to 8 × 106 1 × 104 to 2 × 106 ≤ 13 ml 0.15 MPa, 1.5 bar,21 psi 1.3 ml/min
Sephacryl® S-400 HR(Bulk medium) 2 × 104 to 8 × 106 1 × 104 to 2 × 106 0.5% to 4% of total column volume Column-dependent 10 to 35 cm/h
HiPrep™ 16/60Sephacryl® S-500 HR   4 × 104 to 2 × 107 ≤ 5 ml 0.15 MPa, 1.5 bar,21 psi 0.5 ml/min
HiPrep™ 26/60Sephacryl® S-500 HR   4 × 104 to 2 × 107 ≤ 13 ml 0.15 MPa, 1.5 bar,21 psi 1.3 ml/min
Sephacryl® S-500 HR(Bulk medium)   4 × 104 to 2 × 107 0.5% to 4% of total column volume Column-dependent 10 to 35 cm/h
Sephacryl® S-1000 SF(Superfine,bulk medium)     0.5% to 4% of total column volume not determined 2 to 30 cm/h

* For maximum resolution, apply as small sample volume as possible. Note that sample volumes less than 0.5% normally do not improve resolution.
† See Appendix 5 to convert linear flow (cm/h) to volumetric flow rates (ml/min) and vice versa.

Materials

     
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