Related Experiment Video
Updated: Jul 6, 2026
07:57
Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery
Published on: December 3, 2009
Protein fractionation by preparative electrophoresis
Michael Fountoulakis1, Ploumisti Dimitraki
1F. Hoffmann-La Roche Ltd., Basel, Switzerland.
Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
Summary
Preparative electrophoresis effectively enriches low-abundance proteins using size-based separation in a gel matrix. This technique aids subsequent proteomic analysis for identifying valuable gene products.
Area of Science:
- Biochemistry
- Proteomics
- Molecular Biology
Background:
- Preparative electrophoresis is a key technique for protein fractionation.
- It is commonly utilized in the PrepCell apparatus for isolating specific proteins.
- The method is particularly valuable for enriching low-abundance gene products.
Purpose of the Study:
- To describe the utility of preparative electrophoresis for protein enrichment.
- To highlight its efficiency in isolating low-molecular-mass proteins.
- To outline its integration with subsequent proteomic analyses.
Main Methods:
- Proteins are separated based on size within a cylindrical gel.
- An ionic detergent is used during the separation process.
- The PrepCell apparatus is the standard equipment for this method.
Main Results:
- Achieves efficient enrichment of low-abundance proteins.
- Demonstrates particular efficacy for low-molecular-mass protein targets.
- Eluted proteins are suitable for further detailed analysis.
Conclusions:
- Preparative electrophoresis is a powerful tool for isolating specific protein fractions.
- The technique facilitates subsequent two-dimensional gel electrophoresis and mass spectrometry.
- Enables comprehensive proteomic analysis of enriched low-abundance proteins.
Related Concept Videos
01:22Two-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...
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...
01:32Subcellular Fractionation
The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...
Differential Centrifugation
Differential centrifugation is...
01:20Electrophoresis: 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...
There...
01:27SDS-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...
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...
02:35DNA 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...
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...
01:30Capillary Electrophoresis: Applications
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,...