Strep-tag Probe
Strep-tag Probe - Related Videos
18:27
One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)
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Cited by 12 •
2014
A method is described for efficient purification of twin-Strep-tagged fusion proteins and their specific complexes on modified streptavidin (Strep-Tactin) resin covalently cross-linked with Bis(sulfosuccinimidyl) suberate (BS3). The method has the advantages of fast speed, good target protein recovery and high purity, and is compatible with subsequent analysis by mass spectrometry.
10:59
Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)
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Cited by 9 •
2012
A technique to probe the lipid raft partitioning of fluorescent proteins at the plasma membrane of living cells is described. It takes advantage of the disparity in diffusion times of proteins located inside or outside of lipid rafts. Acquisition can be performed dynamically in control conditions or after drug addition.
08:07
The Microfluidic Probe: Operation and Use for Localized Surface Processing
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Cited by 5 •
2009
In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer solution.
11:26
High-throughput Gene Tagging in Trypanosoma brucei
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Cited by 21 •
2016
Addition of a tag to a protein is a powerful way of gaining insight into its function. Here, we describe a protocol to endogenously tag hundreds of Trypanosoma brucei proteins in parallel such that genome scale tagging is achievable.
05:38
Custom-made Microdialysis Probe Design
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Cited by 9 •
2015
Microdialysis is a commonly used technique in neuroscience research. Therefore commercial probes are in great demand.In this work a probe assembly is explained in detail to build a reliable, concentric, custom-made microdialysis probe for less than $10.
FAQs
Selectivity arises from the engineered interaction between the Strep-tag peptide and Strep-Tactin, an engineered streptavidin derivative. When a probe or capture reagent is linked to a reporter or other functional component, this recognition concentrates that component on the tagged protein rather than distributing it randomly. The result is a targeted signal or retained protein suitable for downstream analysis.
Desthiobiotin acts as a competing ligand that can displace the interaction between Strep-Tactin and the Strep-tag. Because binding can therefore be reversed, researchers can release a captured protein or terminate a detection or isolation step without changing the tag itself. This reversibility adds control to workflows involving purification, pull-down assays, or other capture-based analyses.
Fluorophores support direct visualization, enzymes support signal-generating detection, and other reporter molecules can adapt a probe to the assay format. The underlying Strep-tag interaction remains the targeting step, while the attached reporter determines how the tagged protein is observed or measured. This flexibility lets one recognition strategy support imaging as well as analytical detection.
A practical workflow begins with a recombinant protein carrying the engineered tag, followed by contact with a Strep-Tactin-based probe or capture reagent. The tagged protein is then detected, labeled, or retained for analysis, depending on whether the reagent carries a fluorophore, enzyme, or other reporter. A competing ligand can reverse binding when release is needed.
Application choice depends on the information sought. Protein purification and pull-down assays emphasize recovery or association with other molecules, whereas western blotting emphasizes detection. Fluorescence-based microscopy can reveal cellular localization. Together, these uses allow investigators to examine a recombinant protein’s presence, interactions, distribution, and localization using the same tag-dependent recognition principle.
Tracking where a recombinant protein appears and testing whether it can be captured with associated molecules provides evidence about its distribution and interactions. Those observations can be related to the protein’s function, while selective labeling or isolation helps distinguish the tagged target during analysis. The method therefore connects molecular recognition with cell-level localization and interaction studies.