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Related Concept Videos

Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
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...
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

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...
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...

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Updated: Jun 3, 2026

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells
11:30

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells

Published on: January 26, 2017

The tandem affinity purification technology: an overview.

Yifeng Li1

  • 1Protein Production Core Facility, Department of Biochemistry, University of Texas Health Science Center at San Antonio, 7303 Floyd Curl Drive, San Antonio, TX 78229, USA. liy10@uthscsa.edu

Biotechnology Letters
|March 23, 2011
PubMed
Summary
This summary is machine-generated.

Tandem affinity purification (TAP) isolates protein complexes using a dual-affinity tag for high purity. This method, combined with mass spectrometry, is key for identifying protein interactions in various systems.

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Identification of Protein Interacting Partners Using Tandem Affinity Purification
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2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
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2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Related Experiment Videos

Last Updated: Jun 3, 2026

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells
11:30

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells

Published on: January 26, 2017

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

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
08:23

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein complexes are crucial for cellular functions.
  • Identifying protein interactions is essential for understanding biological processes.
  • Existing purification methods often yield low purity and high background noise.

Purpose of the Study:

  • To provide an overview of the Tandem Affinity Purification (TAP) method.
  • To highlight the significance of the dual-affinity tag in TAP.
  • To discuss the applications of TAP in different biological systems.

Main Methods:

  • Incorporation of a dual-affinity tag into the target protein.
  • Introduction of the tagged protein construct into cells or organisms.
  • Sequential purification using the two affinity handles to isolate protein complexes.
  • Analysis of purified complexes, often using mass spectrometry.

Main Results:

  • TAP enables isolation of protein complexes under physiological conditions.
  • The dual-affinity tag significantly reduces non-specific background.
  • TAP achieves higher purity of isolated protein complexes compared to single-step methods.
  • TAP coupled with mass spectrometry is a standard for identifying multi-protein complexes.

Conclusions:

  • Tandem Affinity Purification is a robust technique for isolating native protein complexes.
  • The dual-affinity tag is the key feature enabling high-purity isolation.
  • TAP is a versatile and widely applicable technology in proteomics research.