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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

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...
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...
Complexometric Titration: Ligands00:43

Complexometric Titration: Ligands

Different monodentate and polydentate ligands are used as complexing agents in complexometric titration reactions. The formation of complexes by mono- and bidentate ligands involves two or more intermediate steps, limiting their use as complexing agents. In comparison, polydentate ligands can form complexes with metal ions in a single-step process, facilitating sharper end points. This means polydentate ligands, such as amino carboxylic acid derivatives, are most commonly employed in...
EDTA: Chemistry and Properties01:22

EDTA: Chemistry and Properties

Polydentate ligands are most widely used in complexometric titrations because they form more stable complexes with the metal ions than mono- or bidentate ligands due to the chelate effect. Examples of polydentate ligands are ethylenediaminetetraacetic acid (EDTA), crown ethers, and cryptands. The most important feature of optimal polydentate ligands is the ability to form 1:1 complexes in a single-step process. Amino carboxylic acid derivatives are frequently used as complexing agents. EDTA is...
Types Of Column Chromatography01:29

Types Of Column Chromatography

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...

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Related Experiment Video

Updated: Jun 3, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
07:53

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

Published on: August 16, 2019

Dye-ligand affinity chromatography.

S J Burton1

  • 1Affinity Chromatography Ltd., Girton, Cambridge, UK.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
PubMed
Summary
This summary is machine-generated.

Dye-ligand affinity chromatography uses a solid support matrix with covalently bonded dye to selectively capture proteins. Bound proteins are then eluted by changing the eluant composition, enabling protein purification.

More Related Videos

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

Related Experiment Videos

Last Updated: Jun 3, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
07:53

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

Published on: August 16, 2019

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

Area of Science:

  • Biochemistry
  • Chromatography
  • Protein Purification

Background:

  • Affinity chromatography is a powerful technique for protein purification.
  • Dye ligands offer versatile binding capabilities for various proteins.

Purpose of the Study:

  • To describe the methodology of dye-ligand affinity chromatography.
  • To highlight the selective binding and elution of proteins using dye-functionalized adsorbents.

Main Methods:

  • Preparation of an adsorbent by covalently bonding a dye to a solid support matrix.
  • Passing a protein mixture through a packed bed of the dye-ligand adsorbent.
  • Washing unbound proteins and eluting bound proteins by modifying eluant composition.

Main Results:

  • Selective binding of proteins capable of interacting with the immobilized dye.
  • Effective separation of target proteins from a complex mixture.
  • Recovery of bound proteins through controlled elution.

Conclusions:

  • Dye-ligand affinity chromatography is an effective method for protein purification.
  • The technique allows for selective capture and subsequent elution of proteins.
  • This method provides a basis for developing specific protein purification strategies.