Homogenisation of protein - (Apr/30/2012 )
Hi, sorry if this a pretty basic topic . 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?
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.