Visualizing Nuclear Pore Complexes in Xenopus Egg Extracts

Sampada Mishra; Daniel L. Levy

doi:10.1007/978-1-0716-2337-4_25

Visualizing Nuclear Pore Complexes in Xenopus Egg Extracts

Sampada Mishra, Daniel L. Levy

Research output: Chapter in Book/Report/Conference proceeding › Chapter

1 Scopus citations

Abstract

The nuclear pore complex (NPC) is the conduit in the nuclear envelope through which proteins and RNA are transported between the cytoplasm and nucleus. Xenopus egg extracts that support de novo assembly of nuclei have provided a robust system to study NPC structure and function because the biochemical composition of the extract can be easily manipulated. Here we describe how to assemble nuclei in Xenopus egg extract, how to visualize and analyze NPCs in both live and fixed samples, and different approaches to altering nucleocytoplasmic transport in extract.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	395-405
Number of pages	11
DOIs	https://doi.org/10.1007/978-1-0716-2337-4_25
State	Published - 2022
Externally published	Yes

Publication series

Name	Methods in Molecular Biology
Volume	2502
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Immunofluorescence
mAb414
NPC
Nuclear import
Nuclear pore complexes
Spindown
Xenopus egg extracts

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10.1007/978-1-0716-2337-4_25

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AU - Mishra, Sampada

AU - Levy, Daniel L.

PY - 2022

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AB - The nuclear pore complex (NPC) is the conduit in the nuclear envelope through which proteins and RNA are transported between the cytoplasm and nucleus. Xenopus egg extracts that support de novo assembly of nuclei have provided a robust system to study NPC structure and function because the biochemical composition of the extract can be easily manipulated. Here we describe how to assemble nuclei in Xenopus egg extract, how to visualize and analyze NPCs in both live and fixed samples, and different approaches to altering nucleocytoplasmic transport in extract.

KW - Immunofluorescence

KW - mAb414

KW - NPC

KW - Nuclear import

KW - Nuclear pore complexes

KW - Spindown

KW - Xenopus egg extracts

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T3 - Methods in Molecular Biology

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BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Visualizing Nuclear Pore Complexes in Xenopus Egg Extracts

Abstract

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