Work

Blue Native Polyacrylamide Gel Electrophoresis (BN‐PAGE) for the Analysis of Protein Oligomers in Plants

Year: 2020

Type: article

Abstract: 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.

Source: Current Protocols in Plant Biology

DOI: 10.1002/cppb.20107

Authors: Pratchaya Pramoj Na Ayutthaya, Derek Lundberg, Detlef Weigel, Lei Li

Institution: Max Planck Institute for Developmental Biology

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: hybrid

Estimated APC paid: $4,750

Funders: Human Frontier Science Program, Deutsche Forschungsgemeinschaft, Max-Planck-Gesellschaft