Figure 3.

HPLC elution profiles of synthetic precursor molecule of GFP after renaturation for 10 days in Tris buffer containing 20% ethylene glycol. (A) RP-HPLC. Column: Pharmacia Resource RPC (1 ml). Eluent: 20–50% CH₃CN in 0.1% TFA. Running conditions: 10 min gradient, 5-min holding; flow rate: 2 ml/min; temp: 25°C. Monitor: Shimadzu Model SPD-M10AVP Photodiode Array UV-VIS Detector attached to the HPLC. mAbsorbance, milli-absorbance unit. (B) Gel-filtration HPLC. Column: Bio-Rad Bio-Select SEC-125–5, 300 × 7.8 mm. Eluent: 0.1 M K₂HPO₄ buffer (pH 7.0). Running conditions: flow rate: 1.0 ml/min; temp: 25°C. Monitor: Same as in A.