The dynamic binding capacity of CM Sepharose™ Fast Flow resin is ~50 mg ribonuclease A/mL resin. Samples were applied at 75 cm/h until 50% breakthrough; columns: 0.5 × 5 cm; buffers: 0.05 M Tris, 2 M NaCl (in the elution buffer), pH 7.5 (DEAE) or 0.1 M acetate, 2 M NaCl (in the elution buffer), pH 5.0. See data file for details.

Buffer pH and ionic strength must be compatible with protein stability and activity. The most suitable pH should allow the proteins of interest to bind, but should be as close to the point of release (elution) as possible. If the pH is too low or too high, elution becomes more difficult and high salt concentrations might be needed. This should be avoided since some proteins begin to precipitate at high ionic strength, and high salt concentrations can interfere with assays or subsequent chromatographic steps. Learn more in Principles and Methods: Ion Exchange Chromatography.

Common troubleshooting topics are discussed in the Instructions for Use and in Principles and Methods: Ion Exchange Chromatography. You can also contact us for support.