4 replies to this topic

[JacobThomas]

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

[bob1]

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

[mdfenko]

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

Edited by mdfenko, 30 November 2012 - 07:25 AM.

[JacobThomas]

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

[mdfenko]

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