Table 1. Different conditions tested for amphipol refolding of BR. Letters in top row represent labels of resulting SEC profiles as shown in Figure 1.
b | c1 | d2 | e | f | g | h | i | j | k | l | m3 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
bacterioOpsin | + | + | + | + | + | + | + | + | + | + | + | + |
Amphipol | - | + | + | + | + | + | + | + | + | + | + | + |
Retinal present | - | + | + | - | - | + | + | + | + | + | + | + |
Bound to Ni-NTA | - | - | - | - | + | - | + | - | + | - | + | - |
KCl-precipitate | + | + | + | + | + | |||||||
Dilute | + | + | ||||||||||
Bio-Beads | + | + |
after Ni-NTA purification with amphipol-free buffer
same as c) but after 12 h incubation with free amphipols
reinjection after first SEC separation (using method in g) and collection of BR containing fraction