Open AccessBlue Native Polyacrylamide Gel Electrophoresis (BN‐PAGE) for the Analysis of Protein Oligomers in Plants
Author(s) -
Na Ayutthaya Pratchaya Pramoj,
Lundberg Derek,
Weigel Detlef,
Li Lei
Publication year - 2020
Publication title -
current protocols in plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.831
H-Index - 7
ISSN - 2379-8068
DOI - 10.1002/cppb.20107
Subject(s) - polyacrylamide gel electrophoresis , polyacrylamide , gel electrophoresis of proteins , color marker , electrophoresis , two dimensional gel electrophoresis , gel electrophoresis , chemistry , chromatography , biochemistry , polymer chemistry , proteomics , enzyme , gene
Protein‐protein interactions, including oligomerization, are involved in regulation of many cellular processes. Unfortunately, many proteins are expressed at a very low level in vivo , making it challenging to observe oligomerization by size‐exclusion chromatography, also known as gel filtration. In this protocol, we present detailed steps to perform blue native polyacrylamide gel electrophoresis (BN‐PAGE), a method to study protein oligomers in plants. The article describes protein sample preparation from transgenic Arabidopsis thaliana and running a BN‐PAGE gel followed by direct western blotting or, alternatively, two‐dimensional sodium dodecyl sulfide–polyacrylamide gel electrophoresis (2D SDS‐PAGE). This protocol will be helpful for new researchers conducting related experiments to analyze stable protein interactions including homo‐ or hetero‐oligomerization in plants. © 2020 The Authors.