Acetic Acid Protein Purfication

Lim Protocol
1. Harvest cells from expression by centrifugation at 18,000 G for 10 minutes at 4C buffer: 10 mM Tris-Cl, 100 mM sodium phosphate, pH = 8 3. Lyse cells with constant cell disrupter at 20 kpsi and centrifuge lysate 4. Collect cell debris, wash in lysis buffer, and re-suspend in 25% (by volume) acetic acid 5. Check purity by SDS-PAGE

2. (See Below) Re-suspend pellet in lysis buffer, 5 mL buffer per gram wet mass. Lysis

Hwang Protocol
1. Harvest cells from expression by centrifugation at 18,000 G for 10 minutes at 4C buffer: 10 mM Tris-Cl, 100 mM sodium phosphate, pH = 8 3. Lyse cells with constant cell disrupter at 20 kpsi and centrifuge lysate (18000 G, 4C, 20 minutes) 4. Collect cell debris and re-suspend in 25% (by volume) acetic acid 5. Centrifuge extracting solution (18000 G, 4C, 20 minutes) 6. Collect supernatant and dialyze in 5% (v/v) acetic acid overnight at 4C

2. (See Below) Re-suspend pellet in lysis buffer, 5 mL buffer per gram wet mass. Lysis

Salt solution prepared by dissolving NaH2PO4 in 5% (v/v), or 0.84 M acetic acid so that final concentration was 0.1 M. pH was adjusted with NaOH.