Tc Tag Specificity
Tc Tag Specificity - Related Videos
10:37
CRISPR/Cas9-mediated Endogenous Fluorescent Tagging of Germline-specific Genes in Caenorhabditis elegans
0 Views •
2026
A CRISPR/Cas9 microinjection workflow for endogenous fluorescent tagging in the Caenorhabditis elegans germline to obtain homozygous knock-in lines for in vivo protein localization analysis.
11:26
High-throughput Gene Tagging in Trypanosoma brucei
0 Views •
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.
10:32
Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
0 Views •
Cited by 7 •
2012
A novel and highly efficient two-step affinity chromatography protocol has been developed and is described in detail. The method is based on a small purification tag with two inherent affinities and is applicable to a wide range of target proteins with different properties.
09:20
Isolation of Subcellular Ribosome Subpopulations Based on Recombinant Peptide Tag-Specific Location-Restricted Illumination-Enhanced Biotinylation
0 Views •
2026
AviTag-specific Location-restricted Illumination-enhanced Biotinylation (ALIBi) is a novel optogenetically activated method developed for labeling and affinity-purifying subcellular populations of ribosomes for downstream sequencing and proteomic analysis.
10:41
Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
0 Views •
Cited by 2 •
2017
A construct encoding TMEM184A with a GFP tag at the carboxy-terminus designed for eukaryotic expression, was employed in assays designed to confirm the identification of TMEM184A as a heparin receptor in vascular cells.
FAQs
Tc tag specificity depends on recognition of the engineered CCXXCC tetracysteine motif by a compatible biarsenical dye. The cysteine-rich sequence provides the binding site that distinguishes the tagged protein from proteins lacking this motif. This selective interaction allows labeling to be directed toward a chosen protein rather than applied broadly across the cell.
FlAsH and ReAsH become fluorescent after binding the tetracysteine tag. Excess unbound dye can then be removed, lowering background fluorescence from molecules that have not attached to the engineered protein. This combination of binding-dependent signal generation and background reduction improves the ability to observe labeled proteins in living cells.
The tetracysteine sequence is relatively small, which may reduce interference with the labeled protein compared with larger fluorescent proteins. Lower structural burden can be valuable when examining protein localization, trafficking, or dynamics, because the tag may be less likely to alter the behavior being measured. The approach therefore supports observation in more native cellular conditions.
A researcher first engineers the tetracysteine sequence into the protein of interest, then uses a biarsenical dye such as FlAsH or ReAsH to bind the introduced motif. Unbound dye is removed before observation to reduce background signal. The resulting fluorescence can be used to follow the tagged protein in living cells.
This labeling strategy can provide information about where a protein is located, how it moves through cellular compartments, and how its distribution changes over time. Because the labeled proteins can be examined in living cells, the method is suited to studying localization, trafficking, and dynamic behavior rather than only fixed endpoint patterns.
Tc tag specificity connects selective molecular labeling with imaging of proteins in their cellular environment. It is relevant to cell biology, molecular imaging, and investigations of protein behavior because fluorescence from the dye-tag complex can make selected proteins visible while unbound dye is reduced. This supports analysis under relatively native biological conditions.