Related Experiment Video
Updated: May 1, 2026
12:03
Blue Native Polyacrylamide Gel Electrophoresis BN-PAGE for Analysis of Multiprotein Complexes from Cellular Lysates
Published on: February 24, 2011
134.7K
One-dimensional SDS-polyacrylamide gel electrophoresis (1D SDS-PAGE)
Julie L Brunelle1, Rachel Green1
1Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Methods in Enzymology
|March 29, 2014
Summary
This protocol details Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), a method to separate proteins by size. SDS-PAGE is crucial for monitoring protein purification, assessing sample purity, and estimating protein molecular weights.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Protein separation is fundamental in molecular biology.
- Accurate assessment of protein purity and size is essential for research and development.
- Established methods are needed for reliable protein analysis.
Purpose of the Study:
- To describe a protocol for Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE).
- To outline the application of SDS-PAGE for protein analysis.
- To provide a method for separating proteins based on molecular weight.
Main Methods:
- Utilizes a denaturing polyacrylamide gel system.
- Employs sodium dodecyl sulfate (SDS) to coat proteins with a uniform negative charge.
- Separates protein molecules based on their size through the gel matrix.
Main Results:
- Proteins are effectively separated according to their molecular size.
- The method allows for clear visualization of distinct protein bands.
- Molecular weights of unknown proteins can be estimated.
Conclusions:
- SDS-PAGE is a versatile and widely applicable technique.
- The protocol enables monitoring of protein purification processes.
- SDS-PAGE is valuable for verifying sample purity and determining molecular weights.
Related Concept Videos
01:27SDS-PAGE
23.2K
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...
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...
23.2K
01:22Two-dimensional Gel Electrophoresis
6.0K
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...
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...
6.0K
01:30Capillary Electrophoresis: Applications
1.9K
Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
1.9K
02:35DNA Agarose Gel Electrophoresis
96.0K
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...
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...
96.0K
01:20Electrophoresis: Overview
4.1K
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...
There...
4.1K
01:15Western Blotting
14.3K
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
14.3K