Protein Purification

RSB250-A Recipe:

• 20mM Tris
• 250mM NaCl
• 30mM Imidazole
• 10% Glycerol (v/v)

• 20mM Tris
• 250mM NaCl
• 500mM Imidazole
• 10% Glycerol (v/v)

Induction

1. Grow a small booster culture in LB Broth overnight (~2-10% of final culture volume) w/ appropriate antibiotics
2. Innoculate full scale culture & monitor OD600 every 15-30 minutes until OD600 reaches 0.8
3. At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter)
4. Allow to grow overnight @ 25°C

Pelleting & Sonication