investigation of protein stability using cyclohexemide - (Oct/12/2006 )

Dear all
I am investigating whether my drug treatment reduces the stability of a certain protein. I just need to check my protocol with you and want to know your opinion about it. I use cycloheximide at a final concentration of 50 microM, I first pre-treat the cells with cycloheximide for 1 hr, then add my drug treatment . I collect samples from different time points and compare (cycloheximide-treated samples) to (cycloheximide plus drug-treated samples).
is this protocol ok?
comments are welcome
thanks

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yup, sounds like what i did... though i was looking at primary/secodary response of drug treatment. i would also include a no cycloheximide treatement control.

v

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I have already included naiive sample (no cycloheximide) and consider this as zero time sample , right??

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you should also perform a parallel time course with the native sample.

then you will have time courses of:

no cycloheximide

cycloheximide

cycloheximide + drug

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