I succesfully did it! You just have to follow the protocol, it is simple! One suggestion: the protein pellet is like gum, I usually use the piston of a syringe to disrupt it and I wash it a lot of times. Moreover, I incubate the pellet for 30' at 37°C to facilitate its resuspension. However, it is almost impossible to totally resuspend it but the quality and the quantity of recovered proteins are good!

Dear John

Here is a protocol provided by Dr. Chomczynski (1987) who invented Trizol Kit:

The organic phase set aside during the DNA isolation step is mixed with 6 ml of isopropanol. The mixture is held at room temperature for ?ve minutes and the precipitated proteins are centrifuged at 10,000 g for ten minutes. The protein pellet is suspended in 3 ml of ethanol by vortexing, then held at room temperature for ?ve

minutes and thereafter centrifuged at 10,000 g for ten minutes. This washing/ centrifuging process is repeated three times. Following the ?nal ethanol wash, the pro tein pellet is brie?y air-dried (ten minutes) and dissolved in water. The protein yield is 6.3 mg. Western analysis are performed according to the standard protocols described in Ausbell, F.M. et al. (Eds): Current Protocols in Molecular Biology,

Wiley Interscience, New York (1990). 50ug of the isolated protein is electrophoresed in 10% acrylamide-SDS gel and electrotransferred to a nitrocellulose membrane.

Good luck,

Thanks very much. We'll try it out and I'll let you know.