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

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...

You might also read

Related Articles

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

Sort by
Same author

Loss of INPP5E affects photoreceptor outer segment membrane biogenesis in iPSC-derived human retinal organoids.

Journal of cell science·2026
Same author

ZDHHC5 interacts physically and functionally with DLG1 at primary cilia and regulates ciliary length and kidney morphology.

Frontiers in cell and developmental biology·2026
Same author

Proximity proteomics reveals a co-evolved LRRK2-regulatory network linked to centrosomes.

EMBO reports·2026
Same author

Organelle contact reorganization drives calcium-dependent autophagy under proteostatic stress.

Autophagy·2026
Same author

Organotypic retinal cultures as an ex vivo screening platform for PLGA-based neuroprotective microspheres.

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences·2026
Same author

Regulation between LRRK2 and PP2A signaling in cellular models of Parkinson's disease.

The Biochemical journal·2026

Related Experiment Video

Updated: Jun 20, 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

Strep/FLAG tandem affinity purification (SF-TAP) to study protein interactions.

Christian Johannes Gloeckner1, Karsten Boldt1,2, Marius Ueffing1,2

  • 1Helmholtz Zentrum München, Neuherberg, Germany.

Current Protocols in Protein Science
|August 19, 2009
PubMed
Summary

A new SF-TAP tag method enables efficient isolation of native proteins and protein complexes. This technique simplifies the analysis of protein-protein interactions in higher eukaryotic cells.

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

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)
18:27

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)

Published on: April 20, 2014

Related Experiment Videos

Last Updated: Jun 20, 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

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)
18:27

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)

Published on: April 20, 2014

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Analyzing protein-protein interactions under native conditions is crucial for understanding cellular processes.
  • Tandem affinity purification (TAP) is a key technique for isolating protein complexes.
  • Existing TAP methods require optimization for efficiency and speed.

Purpose of the Study:

  • To present a novel methodological workflow for rapid and efficient tandem affinity purification.
  • To introduce the SF-TAP tag for isolating native proteins and protein complexes in higher eukaryotic cells.
  • To demonstrate the versatility of SF-TAP for both single protein and complex purification.

Main Methods:

  • Development of an SF-TAP tag, combining Strep-tag II and FLAG moiety.
  • Application of the SF-TAP tag in higher eukaryotic cells for protein purification.
  • Adjustment of purification stringency to achieve desired isolation outcomes.

Main Results:

  • The SF-TAP tag facilitates rapid and efficient tandem affinity purification.
  • The method allows for the isolation of high-purity protein complexes under native conditions.
  • SF-TAP enables selective purification of single tagged-fusion proteins or entire complexes.

Conclusions:

  • The SF-TAP workflow provides an optimized approach for studying protein-protein interactions.
  • This method enhances the ability to analyze native protein complexes in higher eukaryotes.
  • SF-TAP offers a flexible tool for various protein purification strategies.