2.3.1. Protein Immobilization and Hydration
1. Protein: ribonuclease A (RNase, Sigma, Type XII A).
2. Buffer: sodium phosphate 10 mM, pH 7.0.
3. Adsorbing material: celite (Fluka, type 577, particle size <40 |im).
4. Isopiestic equilibration: at a fixed volume ratio of mixtures of water and H2SO4 (minimum 96%, analytical grade). The range of volume ratio may vary from 5 to 20%, according to standard reference tables (26).
5. Labware: a large vacuum-tight dry desiccator or, for very low hydration level, a high vacuum pump; standard Karl-Fisher automatic apparatus for water content determination.
Was this article helpful?