1. Add 222.2 µl 1 M MOPS, pH 7.5 to 2 ml of 10 mg of CREBtide (0.1 M MOPS pH 7.5 final concentration).
2. Read OD205, OD280 (using 0.1 M MOPS buffer as blank).
3. Affigel-10 is stored frozen at < Ð70¡C. Thaw at 4¡C for ~20 min.
4. Filter 1Ð2 ml on nylon (0.22 µM pore size), using Buchner funnel and vacuum. Do not allow beads to dry.
5. Wash bed with 3x2 ml dH2O.
6. Scrape 0.5Ð1 ml into 15 ml microfuge tube.
7. Add 2.22 ml CREBtide ligand solution to tube. Rock at 4¡C for 4 h.
8. Transfer slurry to column.
9. Wash 2x500 µl dH2O.
10. Collect eluate and save at Ð20¡C. Read OD.
11. Wash column:
2x5 ml cold PBS
1x5 ml PBS/0.02% NaN3 (1 µl NaN3 in 5 ml PBS)
12. Store at 4¡C upright in beaker in PBS/NaN3 + parafilm.