Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Affinity Chromatography

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...

Affinity and Avidity

Affinity and Avidity

Electron Affinity

Electron Affinity

The electron affinity (EA) is the energy change for adding an electron to a gaseous atom to form an anion (negative ion).

Molecules and Compounds

Molecules and Compounds

Atoms and Molecules

Chromatography: Introduction

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...

Positive Regulator Molecules

Positive Regulator Molecules

To consistently produce healthy cells, the cell cycle—the process that generates daughter cells—must be precisely regulated.

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Assessment of anti-VEGF intravitreal injection effects on murine neonatal Schlemm's canal morphology.

Scientific reports·2026

Same author

ADAMTS4 elicits myeloid-derived immune cell recruitment and liver fibrogenesis in metabolic dysfunction-associated steatotic liver disease.

Signal transduction and targeted therapy·2026

Same author

A derivative of PD156707 selectively inhibits NLRP3 inflammasome activation by directly binding to NLRP3.

Scientific reports·2026

Same author

Methylated homoisoflavonoids liriopeins A-D as apoptosis inducers and macrophage-activating anticancer agents: Synthesis and biological evaluation.

Bioorganic chemistry·2026

Same author

Sulphur Analogues of Homoisoflavonoids as Potential Treatments for Neovascular Eye Diseases.

ChemMedChem·2026

Same author

Müller Glial Kir4.1 Channel Dysfunction in APOE4-KI Model of Alzheimer's Disease.

Glia·2026

Same journal

Clinical Europium fluorescent based lectin assays for mucin O-glycomics.

Methods in enzymology·2026

Same journal

A dual-color FRET assay for detection and quantitative analysis of O-glycopeptidases.

Methods in enzymology·2026

Same journal

Evolutionary genetic approaches to analyze mucins.

Methods in enzymology·2026

Same journal

Ex vivo imaging and enzymatic analysis of intestinal mucus.

Methods in enzymology·2026

Same journal

Glyco-TRAPP: A real-time glycocalyx permeability assay for assessing transmembrane mucin barrier function in live and fixed tissues.

Methods in enzymology·2026

Same journal

Quantitative imaging approaches to capture structural and functional dynamics of colonic mucus in health and disease in situ.

Methods in enzymology·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jan 24, 2026

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

Small molecule target identification using photo-affinity chromatography.

Seung-Yong Seo¹, Timothy W Corson²

¹College of Pharmacy, Gachon University, Incheon, South Korea.

Methods in Enzymology

|June 4, 2019

Summary

This summary is machine-generated.

Photo-affinity pulldown is a practical method for identifying protein targets of small molecules in chemical biology. This protocol details a versatile approach applicable to various tissues and target identification challenges.

Keywords:

Biotin Click chemistry Inhibitor Mass spectrometry Natural product Off-target Photoreactive group Small molecule Synthetic chemistry Target identification

More Related Videos

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Published on: January 17, 2020

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

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

Published on: January 19, 2022

Related Experiment Videos

Last Updated: Jan 24, 2026

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography

Published on: January 17, 2020

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

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 biology
Drug discovery
Proteomics

Background:

Identifying protein targets of bioactive small molecules is crucial for chemical biology and drug discovery.
Traditional affinity pulldown methods are practical but require specific probe synthesis.

Purpose of the Study:

To provide a detailed protocol for photo-affinity pulldown experiments.
To highlight the general applicability of this technique for target identification.

Main Methods:

Synthesis of affinity probes with crosslinking moieties.
Application of photo-affinity pulldown to diverse tissue types.
Performing experiments with minimal specialized equipment.

Main Results:

Demonstration of a robust photo-affinity pulldown protocol.
Adaptability of the method across various biological samples.
Successful target identification using the described approach.

Conclusions:

Photo-affinity pulldown is a versatile and accessible technique for protein target identification.
The protocol facilitates broad application in chemical biology and drug discovery research.
This method offers a practical solution for identifying bioactive small molecule targets.