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Homogenisation of protein - (Apr/30/2012 )

Hi, sorry if this a pretty basic topic :D. I'm looking to homogenise brain/spinal cord protein, run a non-denaturing western blot, and probe with a labelled drug to look for specific receptors.

1: Can i homogenise to the point of free receptors?
2: What type of homogeniser should i use, glass, polytron or sonication?
3: Will sonication destroy tertiary protein structure and prevent the drug from binding the receptor?

Thanks, Jacob

-JacobThomas-

we always homogenize brain and spinal cord with teflon/glass tissue grinder (and a drill press).

if you use a low clearance homogenizer and an appropriate medium then you should be able to free your receptors.

sonication may denature the proteins.

here are a couple of tissue grinders.

-mdfenko-