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protein induction from yeast


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#1 anwesha1

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Posted 25 September 2012 - 08:39 AM

HI,

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!

#2 ascacioc

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Posted 25 September 2012 - 11:06 AM

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?




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