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Homogenising Brain in 0.32M Sucrose - (Nov/28/2012 )

Hi, just wondering what the reason for homogenising brains in a 0.32M sucrose solution rather than just for example 0.9%PBS?

Thanks, Jacob


Details would help:
Why are you homogenizing brains? what technique are you looking for?


we used .32M sucrose (in buffer) for subcellular fractionation and synaptosome isolation. load on top of sucrose gradient.


Using the brains for a opioid binding studies - crude homogenate, incubate with tritiated opioid + treatment, vacuum filter and look for displacement.

Looking back at some of the binding studies done in the 80's they all seem to homogenise in a sucrose buffer and i can't work out what benefit it would have over PBS.

Thanks, J


the benefit is that it is isoosmotic (like pbs) and ready for application to density gradient centrifugation for fractionation.