Fig. 3

Purification of GFP by alcohol/salt ATPS and HIC. a Successive fractions in the course of alcohol/salt ATPS procedures under UV-illumination. 1, the supernatant of homogenate of infiltrated N. benthamiana leaves after centrifugation; 2, the phase separation showing GFP in the upper ethanol phase after ethanol extraction; 3, the phase separation showing GFP in the lower water phase after addition of n-butanol to the ethanol extract. b Analysis of the various GFP fractions from ATPS procedure by Coomassie-stained polyacrylamide gel. M, protein molecular weight marker in kDa; lanes 1, 2 and 3, the samples corresponding to the fractions from ATPS, as shown in a. c Immunoblot of the various GFP fractions in the course of ATPS. M, protein molecular weight marker in kDa; lanes 1, 2 and 3, the samples corresponding to the fractions in the course of ATPS, as shown in b. d Elution curves of the GFP purified by HIC. e Silver-stained polyacrylamide gel showing the HIC purified GFP. M, Protein molecular weight marker in kDa; lane1, 2, 3 and 4, twofold serial dilution of purified GFP