Preparation of brain lysate - is ultracentrifugation necessary? (Apr/24/2006 )

i want to prepare brain lysate (for the first time in my life ) by this protocol
brain lysate prep

but it says ultracentrifuge....we dont have one here....is it really necessary or I can just centrifuge at max speed available...?? thanx a lot

you can use gxt. the protocol calls for 150000xg for 30-60 min. using the 60 minute spin for the calculation, 60 x 150000 = 9000000. divide 9000000 by the g that you will spin with the slower centrifuge and you will have the time, in minutes, that you will have to spin to equal the clearing efficiency of the ultracentrifugation. be careful, however, the pellet may not be as tight and some components may not spin down as predicted. all in all it should work.

thank you very much for the reply... actually the centrifuge ill be using is in another lab and I am calculating as uve suggested it would mean i would have to centrifuge at least 2 hours at max speed... ...i hope next door lab poeple are ok with that... ...maybe not though...