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Related Concept Videos

Ion Exchange01:17

Ion Exchange

621
Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or...
621
Principles Of Column Chromatography01:13

Principles Of Column Chromatography

6.9K
The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
6.9K
Types Of Column Chromatography01:29

Types Of Column Chromatography

11.2K
The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
11.2K
Chromatography: Introduction01:10

Chromatography: Introduction

4.4K
Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
4.4K
Affinity Chromatography01:03

Affinity Chromatography

731
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
731
Chromatographic Methods: Classification01:12

Chromatographic Methods: Classification

2.4K
Chromatographic techniques are classified in three ways: the classification is based on the physical state of the stationary and mobile phases, how the mobile phase and the stationary phase contact each other, or through the chemical or physical processes that isolate the components of the sample. Typically, the mobile phase is either a liquid or gas, while the stationary phase is either a solid or a liquid layer applied to a solid surface.
Chromatographic techniques are typically named by...
2.4K

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Related Experiment Video

Updated: Jul 17, 2025

Ion Exchange Chromatography IEX Coupled to Multi-angle Light Scattering MALS for Protein Separation and Characterization
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Ion Exchange Chromatography IEX Coupled to Multi-angle Light Scattering MALS for Protein Separation and Characterization

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Ion-Exchange Chromatography: Basic Principles and Application.

Robert G Wallace1, Keith D Rochfort2

  • 1School of Biotechnology, Dublin City University, Glasnevin, Dublin, Ireland.

Methods in Molecular Biology (Clifton, N.J.)
|August 30, 2023
PubMed
Summary
This summary is machine-generated.

Ion-exchange chromatography (IEC) separates molecules using electrostatic properties. This review covers IEC principles and protocols for purifying enzymes from bovine brain tissue.

Keywords:
Anion-exchangeCaptureCation-exchangeContinuous-bedDEAE-Sepharose® Fast FlowLiquid chromatographyPolishProlyl oligopeptidase

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Area of Science:

  • Biochemistry
  • Analytical Chemistry

Background:

  • Ion-exchange chromatography (IEC) is a versatile liquid chromatography technique for separating ionizable molecules based on electrostatic properties.
  • Its wide applicability to proteins and enzymes, coupled with scalability and automation, makes it a cornerstone of biochemical separation.
  • This chapter focuses on the principles and practical application of IEC, specifically anion-exchange chromatography.

Approach:

  • Reviews the fundamental principles and selection criteria for ion-exchange chromatography conditions.
  • Outlines laboratory protocols for the partial purification of a soluble serine peptidase from bovine whole brain tissue.
  • Includes methods for assaying total protein and enzyme activity in fractions before and after purification.

Key Points:

  • IEC separates molecules based on charge differences.
  • Anion-exchange chromatography is demonstrated for enzyme purification from complex biological samples.
  • Detailed protocols cover sample preparation, chromatography, and activity assays.

Conclusions:

  • Ion-exchange chromatography is a powerful and adaptable technique for protein and enzyme purification.
  • The outlined protocols provide a practical guide for researchers purifying enzymes from tissue extracts.
  • Understanding IEC principles is crucial for optimizing separation conditions and achieving successful purification.