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Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE

descriptionPublicationkeyboard_double_arrow_right Article , Other literature type 01 Jan 2018Publisher:Bio-Protocol, LLCJournal:BIO-PROTOCOL, volume 8 (issn: 2331-8325,

Authors: Marjaana Rantala; Virpi Paakkarinen; Eva-Mari Aro;

doi: 10.21769/bioprotoc.2899

pmid: 34395737

pmc: PMC8328655

Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE

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Abstract

The hierarchical composition and interactions of the labile thylakoid protein complexes can be assessed by sequential 2D-native gel-electrophoresis system. Mild non-ionic detergent digitonin is used to solubilize labile protein super-and megacomplexes, which are then separated with first-dimension blue native polyacrylamide gel electrophoresis (1D-BN-PAGE). The digitonin derived protein complexes are further solubilized with stronger detergent, β-DM, and subsequently separated on an orthogonal 2D-BN-PAGE to release smaller protein subcomplexes from the higher-order supercomplexes. Here we describe a detailed method for 2D-BN-PAGE analysis of thylakoid protein complexes from Arabidopsis thaliana.

Related Organizations

University of Turku
Finland

Keywords

QH301-705.5, ta1183, Biology (General)

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