Nucleus and Nuclear Envelope: Methods for Preparation


The nucleus of eukaryotic cells contains the genome surrounded by a nuclear envelope (NE) consisting of two lipid membranes with embedded nuclear pores and an underlying nuclear lamina. The uniformity in size and density makes it possible to isolate pure intact nuclei at high yields from tissue homogenates by centrifugation through a sucrose cushion. NEs can be prepared from isolated nuclei by enzymatic degradation of their nucleic acid content. The resulting NE preparations contain structurally well‐conserved inner and outer nuclear membranes with attached ribosomes, nuclear pore complexes and nuclear lamina. Reliable methods for preparation of nuclei and NEs play an important role in the successful identification of components and their interacting partners that are located in nuclei and in nuclear subcompartments.

Key Concepts

  • Pure intact nuclei can be isolated at high yields from tissue homogenates, and nuclear envelopes can be prepared from the isolated nuclei.
  • The preparation of pure nuclei is important for the identification of nuclear and subnuclear components and their interacting partners.
  • Animal behaviourists must participate in conservation planning to protect the future of biodiversity.
  • Lipid bilayers provide the fundamental architecture of biological membranes.

Keywords: nucleus; nuclear envelope; nuclear membrane; nuclear pore; protein–protein interactions; MCLIP; proteomics

Figure 1. Flowchart showing the major steps from rat liver homogenate to isolated nuclei and isolated nuclear envelopes (NEs).


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Further Reading

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Jafferali, Mohammed H, Beckman, Marie, Kihlmark, Madeleine, and Hallberg, Einar(May 2015) Nucleus and Nuclear Envelope: Methods for Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0002602.pub3]