Table 1.
Solution a | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
---|---|---|---|---|---|---|---|---|---|---|
Tris HCl pH 8.5 (50 mM) | --b | --c | ||||||||
NaCl (150 mM) | --d | |||||||||
GSH/GSSG (5 mM/5 mM) | --e | --f | ||||||||
Cysteine (0.5 mM) | --g | |||||||||
EDTA (5 mM) | ||||||||||
Arginine (0.5 M) | --h | --i | --j | |||||||
% Inhibitionk | 98.5 | 39.6 | 0 | 0 | 98.2 | 93.4 | 92.8 | 97.3 | 87.3 | 98.4 |
Yield (μg/μl)l | 0.05 | <0.01 | <0.01 | <0.01 | 0.03 | 0.04 | 0.03 | 0.03 | 0.04 | 0.03 |
Precipitationm | * | *** | *** | ***** | * | * | *** | ** | ** | ** |
Components used for each solution unless otherwise noted:
Tris HCl, pH 7.4;
MES pH 5;
1 M NaCl;
5 mM/1 mM GSH/GSSG;
1 mM/5 mM GSH/GSSG;
0mM cysteine,
1% CHAPS;
1% sodiumtaurodeoxycholate;
No arginine.
Inhibitory activity of 10 μl of refolded, dialyzed protein on BMP2 signaling in the luciferase reporter assay.
Total protein recovery as measured by BCA.
Precipitation was scored using a five-point scale from one
indicating no precipitation to five
indicating mostly precipitated.