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Homogenising Brain in 0.32M Sucrose


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4 replies to this topic

#1 JacobThomas

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Posted 28 November 2012 - 06:48 PM

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

#2 bob1

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Posted 29 November 2012 - 12:26 PM

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

#3 mdfenko

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Posted 30 November 2012 - 07:25 AM

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.

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#4 JacobThomas

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Posted 10 December 2012 - 03:34 PM

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

#5 mdfenko

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Posted 11 December 2012 - 08:22 AM

the benefit is that it is isoosmotic (like pbs) and ready for application to density gradient centrifugation for fractionation.
talent does what it can
genius does what it must
i do what i get paid to do




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