My protein, a dark, evil mistress from hell, refused to fold on command like her nicely behaving homologous sister. A year into assaying folding and buffer conditions with only slight improvement, I went the mutagenesis route.
After doing all sort of skippy victory dances when I finally got my mutant, I transformed some nice bacteria, grew them and burst them. So I'm, at last, folding the protein again. I expect no problems, I tried folding some of my incomplete mutant and it worked. I dialyzed my protein in the cold room and went home.
I came back this morning and the cold room is at 13 degrees Celsius! Now I have no idea if it folded right, and feel quite guilty of not coming in over the weekend.
I hate my cold room, it's a fickle monster that is constantly under repair. Now I'm squatting in my neighbor's cold room, again, and without a stirring plate :(
Should I precipitate and refold again, to prevent misfolding, or should I just stick it at 4C for the day and hope it folds right?