I am working with isolation of nuclei from human prefrontal cortex.
The protocol I am currently using can be summarized as follows: tissue homogenization using dounce homogenizer followed by a clean-up that is carried out with gradient centrifugation (nuclei suspension is layered on top of a gradient, which is on top of a cushion. After centrifugation the nuclei are collected at the gradient-cushion interphase).
Homogenization buffer, gradient and cushion all contain beta-mercaptoethanol at a concentration of 1 mM.
I would like to substitute beta mercaptoethanol with DTT (Dithiothreitol). I was wondering if anybody using DTT in nuclei isolation could suggest which concentration I should use.