Regeration of Glutathione Sepharose Beads
1, Wash 3 bed volumes of high pH buffer (0.1 M Tris-HCl + 0.5 M NaCl, pH 8.5)
2, Wash 3 bed volumes of low pH buffer (0.1 M sodium acetate + 0.5 M NaCl, pH 4.5)
3, Repeat step 1 & 2 for 3 times.
4, Equilibrate column with 5 bed volumes of PBS.
5, To remove denatured proteins, wash 2 bed volumes of 6 M guanidine hydrochloride, immediately followed by a wash with 5 bed volumes of 1X PBS.
6, To remove hydrophobically bound substances, wash with 4 bed volumes of 70% ethanol or with 2 bed volumes of a non-ionic detergent (conc. 0.1%), immediately followed by a wash with 5 bed volumes of 1X PBS.
