Hi All,

I need to FACS-sort formaldehyde-fixed nuclei from mouse brain tissue. What is the best strategy for staining of intranuclear protein for immunolabeling for FACS (staining of soluble RFP-NLS fusion protein)? Ideally, nuclei will be fixed after or during homogenization of the tissue. I plan to purify nuclei on sucrose cushion and then permeabilize with Tx100. Intracardiac perfusion with PFA is not an option for me (fixation must start after dissection of the brain).

Comments from people who were successful in similar procedures will be highly appreciated.

Cheers,

Lech

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