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protein induction from yeast - (Sep/25/2012 )


My gene is cloned in a galactose-inducible plasmid. I have checked protein expression initially when I checked my plasmids. But after 3 months i'm trying to induce protein expression and do a western blot and i cant find any more protein. i know my clone is correct since i sequenced the stock once more. my galactose solution is about 2.5 months old.. does that matter? i am using the same antibody i had initially used that worked. PLease help!


where did you keep your galactose solution? RT or fridge? I always keep it in fridge because it gets caramelized over time (but not in a few months) I would do a new galactose solution. Any other things you changed/kept too long? Media?