Bio-Rad ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) - Manual (Page 13)

and contaminate the cytoplasmic protein fraction with
nuclear proteins.
Note: Certain tissues may require additional
homogenization to achieve complete cell lysis.
Some optimization may be required.
3. Incubate the Dounce or tissue grinder containing the
homogenate on ice for 1–2 min to allow any large tissue
fragments to sediment to the bottom without pelleting
the nuclei. Using a pipet, carefully transfer the
supernatant to a clean tube, leaving behind any cellular
debris at the bottom of the vessel.
4. Proceed beginning at Step 5 in Section 4.1.1 to
complete the nuclear isolation and solubilization
procedure.
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