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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...
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...
Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatography: Introduction01:10

Chromatography: Introduction

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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Related Experiment Video

Updated: May 27, 2026

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
14:43

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications

Published on: September 23, 2013

Compound immobilization and drug-affinity chromatography.

Uwe Rix1, Manuela Gridling, Giulio Superti-Furga

  • 1CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2011
PubMed
Summary

Identifying the protein targets of bioactive small molecules is crucial for understanding their mechanism of action. This study details a mild chemical proteomics method for drug-affinity chromatography and target identification using mass spectrometry.

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Last Updated: May 27, 2026

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
14:43

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Published on: September 23, 2013

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Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B

Published on: January 19, 2022

Area of Science:

  • Chemical proteomics
  • Drug discovery
  • Molecular pharmacology

Background:

  • Bioactive small molecules can interact with numerous unpredictable protein targets.
  • Defining these targets is essential for understanding a compound's mechanism of action.
  • Chemical proteomics offers a direct approach to identify compound-protein interactions.

Purpose of the Study:

  • To describe a widely applicable procedure for immobilizing small molecule drugs.
  • To outline a method for drug-affinity chromatography (DAC) with subsequent protein identification.
  • To ensure the preservation of molecular integrity during immobilization and chromatography.

Main Methods:

  • Immobilization of small molecule drugs to a solid support using mild conditions.
  • Drug-affinity chromatography to enrich for target proteins.
  • Protein identification via mass spectrometry following enrichment.

Main Results:

  • The described method allows for efficient and mild immobilization of small molecules.
  • Drug-affinity chromatography preserves the integrity of both the compound and target proteins.
  • The procedure is compatible with downstream mass spectrometry analysis for protein identification.

Conclusions:

  • A robust and versatile chemical proteomics protocol is presented for identifying small molecule drug targets.
  • This method facilitates a deeper understanding of drug mechanisms of action.
  • The procedure is crucial for advancing drug discovery and development efforts.