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Updated: Oct 18, 2025
Author Spotlight: Enhancing Drug Discovery - Development of Automated, Standardized Protocols for Nuclei Extraction from Frozen Tissues
Published on: July 28, 2023
NP-40 Fractionation and Nucleic Acid Extraction in Mammalian Cells
Alvaro E Galvis1,2,3,4,5,6, Hugh E Fisher1,2,3,4,5, David Camerini1,2,3,4,5
1Department of Molecular Biology & Biochemistry, University of California, Irvine, CA, USA.
Abstract:
This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear and perinuclear associated components in the nuclear fraction. This protocol has been modified from Sambrook and Russell (2001) in order to downscale the amount of cells needed. To determine the efficiency of fractionation, we recommend using qPCR to compare the subcellular compartments that have been purified with equivalent amount of control whole cell extracts.
