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Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

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...
Capillary Electrophoresis: Applications01:30

Capillary 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,...
Electrophoresis: Overview01:20

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...
Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
SDS-PAGE01:27

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...

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Related Experiment Video

Updated: Jun 6, 2026

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids
10:32

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids

Published on: March 2, 2012

Isoelectric focusing in a drop.

Noah G Weiss1, Mark A Hayes, Antonio A Garcia

  • 1Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287-9709, United States.

Langmuir : the ACS Journal of Surfaces and Colloids
|December 2, 2010
PubMed
Summary

Droplet-based isoelectric focusing enables molecular separations within discrete drops on a superhydrophobic surface. This novel technique achieves preparative separation by manipulating and splitting droplets after pH gradient generation.

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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

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Separation of Bioactive Small Molecules, Peptides from Natural Sources and Proteins from Microbes by Preparative Isoelectric Focusing (IEF) Method
09:57

Separation of Bioactive Small Molecules, Peptides from Natural Sources and Proteins from Microbes by Preparative Isoelectric Focusing (IEF) Method

Published on: June 14, 2020

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Last Updated: Jun 6, 2026

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids
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Published on: March 2, 2012

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
07:58

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

Separation of Bioactive Small Molecules, Peptides from Natural Sources and Proteins from Microbes by Preparative Isoelectric Focusing (IEF) Method
09:57

Separation of Bioactive Small Molecules, Peptides from Natural Sources and Proteins from Microbes by Preparative Isoelectric Focusing (IEF) Method

Published on: June 14, 2020

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Microfluidics

Background:

  • Isoelectric focusing is a key protein separation technique.
  • Current methods can be complex and require specialized equipment.
  • Need for simplified, high-throughput separation methods.

Purpose of the Study:

  • Investigate droplet-based isoelectric focusing for molecular separations.
  • Develop a novel, discrete droplet manipulation technique for preparative separation.
  • Validate the efficiency and applicability of the new method.

Main Methods:

  • Utilized droplet manipulation on a superhydrophobic surface.
  • Applied low voltages for isoelectric focusing within discrete drops.
  • Generated stable, reversible pH gradients (3-10) using ampholyte buffers.
  • Split droplets to achieve preparative separation.
  • Characterized protein focusing via visual, spectroscopic, and light scattering methods.

Main Results:

  • Demonstrated stable, reversible pH gradients (3-10) using a universal indicator dye.
  • Achieved protein focusing within the length of individual drops.
  • Quantitatively verified focusing using noninvasive light scattering.
  • Results correlated well with a 1D steady-state theoretical model.
  • Successfully separated differential fractions into new discrete liquid elements.

Conclusions:

  • Droplet-based isoelectric focusing is a viable novel approach for molecular separations.
  • Preparative separation is achievable within a single, open droplet.
  • This technique offers a simplified and potentially high-throughput method for biochemical analysis.