Hello everyone,

First time for me here, in my PhD, I would like to expose intact and functional nuclei to a compound for a few hours at 37 degrees.

We already use the Abcam Nuclear Fractionation Protocol. I was wondering if my nuclei were still functional and intact after adding the hypotonic lysis buffer. Here is the composition of buffer A from Abcam:

Buffer A: 10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT, 0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9 To prepare 250 ml stock of buffer A – HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250ml MgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250ml KCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250ml DTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250ml NP40 = 0.05%

Or it is better for me to buy nuclear isolation kit ?

With your experiences if you have any suggestions, I am listening to you, thank you very much

Charlotte