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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

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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...
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Principles Of Column Chromatography01:13

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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...
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Chromatography: Introduction01:10

Chromatography: Introduction

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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...
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Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

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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...
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Types Of Column Chromatography01:29

Types Of Column Chromatography

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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
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Silica Gel Column Chromatography: Overview01:10

Silica Gel Column Chromatography: Overview

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Silica gel column chromatography is a technique for separating compounds using a column packed with silica gel as the stationary phase. This method relies on differences in the polarity of compounds. Based on their polarities, compounds move between the stationary phase (silica gel) and the mobile phase (the solvent), forming discrete bands in the column.
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Related Experiment Video

Updated: Apr 25, 2026

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
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Frontal affinity chromatography (FAC): theory and basic aspects.

Ken-ichi Kasai1

  • 1Department of Biological Chemistry, School of Pharmaceutical Sciences, Teikyo University, 2-11-1 Kaga, Itabashi, Tokyo, 173-8605, Japan, kasai-k-4649@hotmail.co.jp.

Methods in Molecular Biology (Clifton, N.J.)
|August 14, 2014
PubMed
Summary
This summary is machine-generated.

Frontal affinity chromatography (FAC) is a powerful technique for studying biomolecule interactions. This method is especially valuable in glycobiology for understanding molecular binding.", Enhanced_Abstract=default_api.SeocontentEnhancedAbstract(Area_of_Science=[

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Area of Science:

  • Biochemistry and Molecular Biology
  • Analytical Chemistry
  • Glycobiology

Background:

  • Understanding specific interactions between biomolecules is crucial in biological research.
  • Glycobiology, the study of carbohydrates, relies heavily on methods to analyze carbohydrate-protein interactions.

Purpose of the Study:

  • To present the fundamental principles, advantages, and theoretical basis of Frontal Affinity Chromatography (FAC).
  • To highlight the utility of FAC as an analytical tool in biomolecular interaction studies.

Main Methods:

  • Detailed explanation of the theoretical underpinnings of Frontal Affinity Chromatography.
  • Discussion of the practical aspects and setup of FAC experiments.
  • Exploration of data analysis methods specific to FAC.

Main Results:

  • Demonstration of FAC's capability to determine binding constants and stoichiometry.
  • Illustrative examples of FAC application in analyzing complex biological systems.
  • Highlighting the versatility and robustness of the FAC technique.

Conclusions:

  • Frontal Affinity Chromatography is a highly effective method for quantitative analysis of biomolecular interactions.
  • FAC offers significant advantages for researchers in glycobiology and other fields requiring precise interaction analysis.
  • The presented overview provides a foundational understanding for implementing and interpreting FAC studies.