These rigid macroporous hydrophilic media meet the demands of process-scale applications. Chemically and mechanically stable, the media operate well at low and medium pressures.
Dynamic protein binding capacity as a function of linear flow rate. A 100 ml sample (3 mg/ml) of BSA was run on a 1 x 10 cm (8 ml) column at the indicated flow rates. The buffer was 50 mM Tris-HCI, pH 8.3. Elution was with 1.0 M NaCl.
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The Macro-Prep DEAE derivative has high dynamic binding capacity and exhibits consistently high resolution even at high flow rates and high sample loads. A sample containing 3 mg/ml BSA was separated on a 1 x 10 (3 ml) Macro-Prep DEAE column. Sample was loaded onto the column in 10 mM Tris-HCI, pH 7.6 (buffer A) and was eluted in a M NaCl in buffer A. Sample was applied until the eluting buffer had an A280 value 50% of the incoming sample.
The effect of flow rate on separation and peak symmetry. A 5 ml sample containing 3.65 mg/ml each myoglobin, conalbumin, BSA, and soybean trypsin inhibitor was loaded onto a column containing 6.56 ml Macro-Prep DEAE support. The sample was loaded in 50 mM Tris-HCl, pH 7.6 (buffer A) and eluted in a gradient of 0–1 M NaCl in buffer A as follows: buffer A for 15 ml, 0–0.35 M NaCl in 85 ml, 0.35–0.65 M NaCl in 15 ml, 0.65–1 M NaCl in 15 ml.
Macro-Prep High Q media: Effect of flow rate on peak symmetry and separation. A 5 ml sample (4.4 mg/ml) of myoglobin (peak 1), ovalbumin (peak 2), and BSA (peak 3) was run on a 1 x 13 cm (8.7 ml) column at each of the indicated flow rates. The buffer was 50 mM Tris-HCl, pH 8.6, with a gradient of 0–0.5 M NaCl over 100 ml.
Ammonium sulfate fraction of Hevea (rubber plant) cytosol on Macro-Prep 25 Q (11 x 21 cm). Buffer A, 50 mM Tris, pH 8.3; buffer B, 50 mM Tris, 1 M NaCl, pH 8.3; flow rate = 5 ml/min.
Separation on 360µl of 6x His-tagged dihydrofolate reductase E. coli lysate on a 2 ml column packed with Macro-Prep 25 Q Media. Buffer A: 50 mM Tris, pH 8.3. Buffer B: 50 mM Tris, pH 8.3, 1 M NaCl; gradient: 0–100% B in 10 column volumes; flow rate: 2 ml/min.
Sample separation with the Macro-Prep DEAE support and two other commercially available DEAE supports. A 100 µl sample containing 9 mg/ml each of myoglobin, conalbumin, ovalbumin, BSA, and soybean trypsin inhibitor was loaded onto a 0.5 cm ID column containing 2 ml of support. Sample was loaded onto the column in 50 mM Tris-HCl, pH 7.6 (buffer A) and was eluted with 1 M NaCl in buffer A. The flow rate was 153 cm/hr (0.5 ml/min). Scales offset for comparison.
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