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TCA protein precipitation - (Feb/03/2005 )

We've been using TCA precipitation to prepare samples for western blot. However, some proteins can not been detected using this method. For example, nuclear protein lamin-B (55kDa). Anybody knows why?


TCA is bad, don't use it. It is inconsistent and relies on several factors that not all proteins have. It also causes problems with the gels.
Better method to precipitate proteins. wink.gif


i got good result with TCA precipitation
here is the recipe :

mix sample 100% ice cold acetone (sotred at -20 and keeped on ice during exp) and 100%TCA in 1:8:1 ratio
incubation -20° 1h
spin 18000g 15' 4°
wah with 1vol 100% cold acetone
spin 18000g 15' 4°

wash with ethanol twice (1ml each) and let air dry for 30'
add rehycratation buffer and let sit 1h vortexing every 10'
Transfert to new eppendorf tube and spin max speed 10' RT
Transfert immediately to new eppy and store at -80°
ready to use