Solubilization buffer, DIGE
10% (v/v) glycerol
30 mM HEPES
150 mM potassium acetate
Adjust the pH to 8.0 with KOH. Add the detergent of choice:
Heat buffers containing digitonin to >95°C in a heating block to facilitate complete dispersion. Once in solution, digitonin does not precipitate for a few hours, even if it is cooled down to 4°C.
n-Dodecyl maltoside
The choice of detergent used (and its concentration) should be determined empirically for a given system before commencing the study. For respiratory complexes, 5% (w/v) digitonin is recommended. Other samples might be better served using 2%-5% digitonin, 0.1%-2% (v/v) Triton X-100, or 0.5%-2% (w/v) n-dodecyl maltoside.
