protease inhibitor, TRI reagent and western - (Oct/23/2008 )

Hi
Is it necessary to add protease inhibitor when isolating protein for western blot using TRI reagent (from cell culture)?
If so, when?
Thanks

HI,

I recommend to use protease inhibitors also for trizol purification. i would add it right at the beginning as the inhibitor binds to the protease, stays with it and protects your proteins from degradation. we usually use the convenient " complete " inhib. tablets from roche.

best wishes
steph

Thanks for that,
Did you mean the inhibitor should be added at the very beginning of the protocol or at the beginning of the protein isolation section?

Thanks again