I would like to measure glucocorticoid receptor translocation and phosporylated isoforms of this receptor in both fractions of PVN and ventral hippocampus. I would appreciate any advices since I have not tried this before. Are there any specialties to be aware of?
If you have some tissue or cell cluster you can use the kit what we used to see cytosolic and nuclear proteins: http://jvi.asm.org/content/early/2013/06/05/JVI.03385-12.abstract
is it suitable for frozen tissues as well? I am just affraid most commercial kits are not sufficient for special purposes, like frozen tissues of small size.
I do agree with your concern, you may be requiring a strong buffer to solubilise the tissue first and then fractionate different compartments with those selective buffers. There may be some kits available for this purpose. But as a start, why dont you just homogenize the tissue and try using some kit/appropriate method to test what happens...good luck!
:-) the reason is very simple, not enough time, but anyway, I got few methods how to isolate it but still would like to consult with someone who is really experienced and could provide me some details which are perhaps not available elsewhere. Yep, definitely I will try something.
I am also interested in finding a good protocol for this purpose. We are working with small, frozen rat brain tissue, and require processing a couple hundred samples at a time, and thus the commercial kits are just not an option due to price.
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