I would like to use 100% methanol (-20 degree C) for fixing monolayer cell culture for ICC-type of procedures. Is there any requirement on the grade of methanol to be used? E.g. Sigma has this (Cat. No. 494437) methanol "Suitable for protein sequencing, BioReagent" and this (Cat. No. M1775) listed under "fixatives", while we have analytical grade methanol (for preparing Western blot transfer buffer) in the lab. Can someone advice which grade of methanol should I use? Many thanks!
normal clean methanol is sufficient. I think we use bioreagent grade. Be careful to use really cold methanol and add it slowly and dropwise if possible drip it on the side of your well because you cann easily detach cells by methanol addition in case it is to harsh
Analytical grade methanol should be fine. I'm curious to know, why do you keep methanol @ -20 C? generally methanol stored at 4 C is used for fixing?
Thanks guys for your answers and advice!
As for why keeping methanol at -20 C, hmmm... that's what I was taught to do; I seem to have the impression that since methanol is a very powerful solvent of the membrane, using it at -20 would slow down its action and prevent it from acting too harshly on the cells. Please correct me if that's not the right reason!
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