Self-labeling Tags
Self-labeling Tags - Related Videos
09:06
Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
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Cited by 10 •
2017
Combined precursor isotopic labeling and isobaric tagging (cPILOT) is a quantitative proteomics strategy that enhances sample multiplexing capabilities of isobaric tags. This protocol describes the application of cPILOT to tissues from an Alzheimer's disease mouse model and wild-type controls.
07:38
Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
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Cited by 8 •
2010
SNAP-tag and CLIP-tag protein labeling systems enable the specific, covalent attachment of molecules, including fluorescent dyes, to a protein of interest in live cells. Once cloned and expressed, the tagged protein can be used with a variety of substrates for numerous downstream applications without having to clone again.
09:24
Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
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Cited by 2 •
2020
Combined precursor isotopic labeling and isobaric tagging (cPILOT) is an enhanced sample multiplexing strategy that is capable of increasing the number of samples that can be analyzed simultaneously with available isobaric tags. Incorporation of a robotic platform has greatly increased experimental throughput, reproducibility, and quantitative accuracy.
08:28
Metabolic Labeling
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2023
Metabolic labeling is used to probe the biochemical transformations and modifications that occur in a cell. This is accomplished by using chemical analogs that mimic the structure of natural biomolecules. Cells utilize analogs in their endogenous biochemical processes, producing compounds that are labeled. The label allows for the incorporation of detection and affinity tags, which can then be used to elucidate metabolic pathways using other biochemical analytical techniques, such as SDS-PAGE...
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.
FAQs
The engineered enzyme component recognizes a compatible synthetic substrate, such as benzylguanine or benzylcytosine, and then forms a covalent bond with it. When the substrate carries a fluorescent dye or another probe, that reaction attaches the signal directly to the tagged protein, supporting selective observation in living cells.
Because the tag-substrate connection is stable, the attached probe remains associated with the protein during the observation period. This persistence helps researchers follow protein localization or movement without relying on a transient interaction. It also supports turnover measurements, where labeling at selected times can distinguish changes in the protein population over time.
Using different substrates at different time points gives the experiment temporal flexibility. One probe can mark a protein at an earlier stage, while another probe can be introduced later, allowing sequential observations of the same tagged system. This design is particularly useful for pulse-chase experiments and for examining protein trafficking or turnover.
A typical live-cell workflow begins by genetically encoding the tag with the protein of interest, followed by exposure to a compatible synthetic substrate linked to a fluorescent dye or other probe. After the covalent labeling reaction, researchers examine the protein's localization, movement, or turnover under the chosen observation conditions.
Pulse-chase experiments are useful when timing matters more than a single endpoint. Researchers can label proteins at selected times and then use later labeling or observation to assess how those proteins move through the cell or change over time. The approach therefore connects molecular labeling with dynamic studies of trafficking and turnover.
Self-labeling Tags can be selected when a study needs information about proteins in complex biological systems rather than only a fixed snapshot. Live-cell imaging reveals localization and movement, while the same strategy can support studies of turnover, interactions, or biochemical purification. Its value lies in linking timed molecular labeling to several protein-level readouts.