May 29, 2018 · Prior to binding, beads were washed twice with one volume (equal to the initial bead volume) of Buffer A (100 mM NaOH, 50 mM NaCl), once with one volume of Buffer B (100 mM NaCl), and once with one volume of low-salt binding Buffer C (20 mM Tris pH 8.0, 140 mM KCl, 5 mM MgCl2, 100 μg/mL CHX, 0.5 mM DTT, 0.1% Triton X-100).