Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Two-dimensional Gel Electrophoresis

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...

SDS-PAGE

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...

DNA Agarose Gel Electrophoresis

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...

Electrophoresis: Overview

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Kidins220 regulates the development of B cells bearing the λ light chain.

eLife·2024

Same author

The role of the different CD3γ domains in TCR expression and signaling.

Frontiers in immunology·2022

Same author

Wiskott-Aldrich Syndrome Protein: Roles in Signal Transduction in T Cells.

Frontiers in cell and developmental biology·2021

Same author

Spatiotemporally confined red light-controlled gene delivery at single-cell resolution using adeno-associated viral vectors.

Science advances·2021

Same author

Cross-TCR Antagonism Revealed by Optogenetically Tuning the Half-Life of the TCR Ligand Binding.

International journal of molecular sciences·2021

Same author

Cooperative Interaction of Nck and Lck Orchestrates Optimal TCR Signaling.

Cells·2021

Search research articles

Related Experiment Video

Updated: Jul 6, 2026

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Published on: February 24, 2011

Two-dimensional blue native polyacrylamide gel electrophoresis.

Wolfgang W A Schamel¹

¹Max Planck-Institut für Immunbiologie und Universität Freiburg, Biologie III, Freiburg, Germany.

Current Protocols in Cell Biology

|March 25, 2008

Summary

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) is a versatile technique for analyzing protein complexes. This study provides detailed protocols for two-dimensional BN/SDS-PAGE and stoichiometry determination.

More Related Videos

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Published on: September 28, 2018

Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

Published on: February 12, 2019

Related Experiment Videos

Last Updated: Jul 6, 2026

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Published on: February 24, 2011

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Published on: September 28, 2018

Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

Published on: February 12, 2019

Area of Science:

Cell Biology
Biochemistry
Proteomics

Background:

Multiprotein complexes are essential for cellular functions.
Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) is a key technique for studying these complexes.
BN-PAGE offers higher resolution than traditional methods like gel filtration.

Purpose of the Study:

To provide detailed protocols for two-dimensional BN/SDS-PAGE.
To present a related technique for determining multiprotein complex stoichiometry.
To facilitate the study of protein-protein interactions and complex composition.

Main Methods:

Two-dimensional Blue Native Polyacrylamide Gel Electrophoresis (BN/SDS-PAGE) for separating protein complexes.
Coomassie blue dye is used to impart charge for native gel electrophoresis.
A complementary method for assessing the stoichiometry of protein complexes.

Main Results:

Detailed protocols are provided for BN/SDS-PAGE and stoichiometry analysis.
The methods allow for the separation and characterization of protein complexes ranging from 10 kDa to 10 MDa.
The study supports the increasing use of BN-PAGE in protein interaction research.

Conclusions:

Two-dimensional BN/SDS-PAGE is a powerful tool for analyzing multiprotein complex size, subunit composition, and abundance.
The described protocols enhance the capability to study complex stoichiometry.
These methods are valuable for advancing research in cell biology and proteomics.