Blue Native Page
Blue Native Page - Related Videos
12:03
Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
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Cited by 92 •
2011
In this video, we describe the characterization of multiprotein complexes (MPCs) by blue native polyacrylamide gel electrophoresis (BN-PAGE). In a first dimension, dialyzed cellular lysates are separated by BN-PAGE to identify individual MPCs. In a second dimension SDS-PAGE, MPCs of interest are further subdivided to analyze their constituents by immunoblotting.
09:35
Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
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Cited by 9 •
2017
Here we present protocols for affinity purification of protein complexes and their separation by blue native PAGE, followed by protein correlation profiling using label free quantitative mass spectrometry. This method is useful to resolve interactomes into distinct protein complexes.
06:59
Multimer-PAGE for Separating Native Protein Complexes: A Hybrid Separation Technique Consisting of Blue Native-PAGE and SDS-PAGE to Separate Intact Multimeric Proteins From Tissue Lysate
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2025
This video describes multimeric PAGE for separating native protein complexes from tissue homogenates. The technique is a hybrid of blue native-PAGE and SDS-PAGE techniques. The separated complexes can be characterized and studied for their role in cell functioning.
03:10
Blue Native Polyacrylamide Gel Electrophoresis: A Non-Denaturing Separation Technique for Analysis of Intact Mitochondrial Respiratory Chain Complexes
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2025
In this video, we demonstrate an electrophoretic technique, called blue native PAGE, for the analysis of mitochondrial respiratory protein complexes in their natively folded and functionally active state. The anionic Coomassie blue dye causes the protein complexes to become negatively charged, allowing separation based on size and structure.
08:12
Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis
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Cited by 15 •
2018
A protocol for the elucidation of plant thylakoid protein complex organization and composition with blue native polyacrylamide gel electrophoresis (BN-PAGE) and 2D-SDS-PAGE is described. The protocol is optimized for Arabidopsis thaliana, but can be used for other plant species with minor modifications.
FAQs
Coomassie Brilliant Blue binds to proteins and supplies a negative charge without requiring complete complex dissociation. This added charge allows intact assemblies to move through the polyacrylamide gel during electrophoresis. Because the complexes retain native organization, their migration reflects the combined influence of size, shape, and charge rather than charge alone.
Migration depends on the complex's size, shape, and charge. These properties act together, so gel position does not provide a simple molecular-weight measurement comparable to a fully denaturing separation. Interpreting a band therefore requires considering how the assembly's overall structure and Coomassie-derived charge influence its movement through the gel.
Blue Native Page maintains many native interactions and activities, whereas SDS-based conditions can fully dissociate protein assemblies. This distinction makes the native method suitable for observing intact oligomers, respiratory-chain complexes, and supercomplexes. SDS-PAGE remains useful afterward because a second dimension can separate the components released from those assemblies and reveal their composition.
A typical workflow separates the protein preparation through a polyacrylamide gel under non-denaturing conditions, using Coomassie Brilliant Blue to provide electrophoretic charge. The resulting bands represent assemblies whose native organization has been retained to a substantial extent. Researchers can then evaluate their composition with second-dimension SDS-PAGE or examine activity directly within the gel.
In-gel activity assays help determine whether separated complexes retain detectable function after electrophoresis. Because Blue Native Page preserves many native interactions and activities, a band can be assessed not only by its migration position but also by its activity signal. This connects the observed assembly with functional information, especially when studying membrane or respiratory-chain complexes.
The technique can reveal whether proteins occur as individual assemblies, oligomeric states, or larger supercomplexes. In membrane biology and respiratory-chain research, this supports analysis of how complexes are organized while preserving many native associations. Pairing the separation with activity assays or a second SDS-PAGE dimension adds functional or compositional context to the observed bands.