Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

3.5K
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...
3.5K
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

8.7K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
8.7K
Immunoprecipitation01:20

Immunoprecipitation

8.0K
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...
8.0K

You might also read

Related Articles

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

Sort by
Same author

Calcium/calmodulin-dependent protein kinase II links ER stress with Fas and mitochondrial apoptosis pathways.

The Journal of clinical investigation·2009
Same author

Cripto-1 overexpression is involved in the tumorigenesis of nasopharyngeal carcinoma.

BMC cancer·2009
Same author

Range of motion and orientation of the lumbar facet joints in vivo.

Spine·2009
Same author

[Silencing of COX-2 in nasopharyngeal carcinoma cells with a shRNAmir lentivirus vector].

Nan fang yi ke da xue xue bao = Journal of Southern Medical University·2009
Same author

The risk of melamine-induced nephrolithiasis in young children starts at a lower intake level than recommended by the WHO.

Pediatric nephrology (Berlin, Germany)·2009
Same author

Adult scoliosis in patients over sixty-five years of age: outcomes of operative versus nonoperative treatment at a minimum two-year follow-up.

Spine·2009

Related Experiment Video

Updated: Apr 5, 2026

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

4.3K

Affinity purification in target identification: the specificity challenge.

Wenlu Zheng1, Gang Li2, Xiaoyu Li3,4

  • 1Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, 518055, China.

Archives of Pharmacal Research
|August 8, 2015
PubMed
Summary
This summary is machine-generated.

Phenotype-based screening identifies potential drugs, but target identification is crucial. This review discusses affinity purification methods to overcome challenges in finding specific small molecule targets within complex biological systems.

Keywords:
Affinity purificationPhenotypic screeningProtein–ligand interactionSmall moleculeSpecificityTarget identification

More Related Videos

Identification of Protein Interacting Partners Using Tandem Affinity Purification
10:02

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Published on: February 25, 2012

38.4K
Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

14.7K

Related Experiment Videos

Last Updated: Apr 5, 2026

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

4.3K
Identification of Protein Interacting Partners Using Tandem Affinity Purification
10:02

Identification of Protein Interacting Partners Using Tandem Affinity Purification

Published on: February 25, 2012

38.4K
Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

14.7K

Area of Science:

  • Pharmaceutical Sciences
  • Chemical Biology
  • Molecular Pharmacology

Background:

  • Phenotype-based screening is vital for drug discovery, evaluating small molecule bioactivity in biological systems.
  • Identifying molecular targets is essential for understanding drug mechanisms and optimizing therapeutic potential.
  • Affinity purification is a key strategy for revealing direct physical interactions between small molecules and their targets.

Purpose of the Study:

  • To review the history and background of affinity purification in target identification.
  • To discuss technological advancements addressing the specificity challenge in affinity purification.
  • To categorize and highlight methods for distinguishing true targets from non-specific interactions.

Main Methods:

  • Review of historical and recent developments in affinity purification techniques.
  • Categorization of methods into noise reduction and comparative distinction strategies.
  • Emphasis on integrated approaches combining multiple techniques for robust target identification.

Main Results:

  • Affinity purification is powerful but faces specificity challenges due to proteome complexity.
  • Technological developments aim to enhance target identification accuracy by reducing background noise.
  • Comparative distinction methods are crucial for differentiating specific interactions from non-specific binding.

Conclusions:

  • Overcoming specificity challenges in affinity purification is critical for successful target identification.
  • A combination of noise reduction and comparative distinction methods offers a robust strategy.
  • Integrated approaches are highlighted as essential for achieving reliable identification of small molecule targets.