Epitope Tag Reporters
Epitope Tag Reporters - Related Videos
13:58
Peptide:MHC Tetramer-based Enrichment of Epitope-specific T cells
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Cited by 28 •
2012
This protocol describes the use of peptide:MHC tetramers and magnetic microbeads to isolate low frequency populations of epitope-specific T cells and analyze them by flow cytometry. This method enables the direct study of endogenous T cell populations of interest from in vivo experimental systems.
07:16
Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions
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Cited by 1 •
2024
Protein-protein interactions are important for elucidating the function of target proteins, and co-immunoprecipitation (co-IP) can easily confirm PPIs. We transiently transfected a plasmid encoding an epitope-tagged protein into HEK-293 cells and developed an immunoprecipitation method to easily confirm the binding of two target proteins.
08:07
Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
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2024
Here, we describe a newly developed multiplex fluorescent immunoassay that uses a dual-reporter flow cytometric system to concurrently detect two unique spike protein epitopes on intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles that had been captured by angiotensin-converting enzyme-2-coupled magnetic microspheres.
08:10
A Functional Whole Blood Assay to Measure Viability of Mycobacteria, using Reporter-Gene Tagged BCG or M.Tb (BCG lux/M.Tb lux)
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Cited by 19 •
2011
We describe an alternative approach to the enumeration of mycobacteria in vitro, which uses reporter-gene tagged mycobacteria instead of colony-forming units (CFU). “Survival” of organisms as well as host response-markers are measured simultaneously, providing a low-cost, versatile and functional system for studies of host/pathogen interactions in the context of tuberculosis.
05:45
In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin
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Cited by 4 •
2019
Morphological changes occur in immune responsive fibroblast cells following activation and promote alterations in cellular recruitment. Utilizing 2-photon imaging in conjunction with a genetically engineered Fibroblast-specific protein 1 (FSP1)-cre; tdTomato floxed-stop-floxed (TB/TB) mouse line and green fluorescently tagged lipopolysaccharide-FITC, we can illustrate highly specific uptake of lipopolysaccharide in dermal fibroblasts and morphological changes in vivo.
FAQs
Detection begins when the short peptide epitope is genetically fused to the protein of interest. The resulting engineered protein carries a recognizable molecular feature while retaining the identity of the target protein. Tag-specific antibodies can then bind that feature and reveal where the protein is located or how much is present, even when a suitable antibody against the native protein is unavailable.
A standardized epitope provides a consistent recognition site for tag-specific antibodies across samples. This can make differences in detected signal easier to compare when researchers examine protein expression under different experimental conditions. In behavior research, such comparisons help associate molecular changes in neuronal proteins with changes observed between behavioral groups or treatments.
The detection method determines the type of information obtained. Immunostaining reveals the cellular location of the tagged protein, allowing researchers to examine where it appears in neuronal tissue or cells. Immunoblotting supports assessment of protein abundance in a sample. Using either approach, or interpreting their results together, connects protein distribution or level with experimental outcomes.
They are particularly useful when an endogenous antibody is unavailable or does not provide sufficient specificity. Instead of depending on an antibody that recognizes the native protein reliably, researchers can detect the engineered epitope with a tag-specific antibody. This creates a more standardized route for examining target proteins and supports consistent analysis across behavioral experiments.
A typical application starts by genetically fusing the short epitope to the selected protein of interest. Researchers then examine the engineered protein using tag-specific antibody detection, commonly through immunostaining or immunoblotting. The resulting measurements can be compared across experimental conditions and related to behavioral outcomes, depending on whether the study emphasizes cellular location or protein abundance.
Researchers can measure a tagged neuronal protein after subjects or samples have been examined under defined behavioral conditions, then compare its abundance or cellular location across those conditions. This provides a molecular readout that can be interpreted alongside behavioral results. The approach helps investigate whether changes in protein expression or localization accompany differences in behavior.
These reporters can provide two complementary forms of information: the relative abundance of a target protein and its cellular location. Immunoblotting is suited to examining abundance, whereas immunostaining shows distribution within cells or tissue. In behavior research, those measurements help characterize molecular changes in neuronal proteins and determine how they relate to behavioral findings.