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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

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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...
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Related Experiment Video

Updated: Dec 21, 2025

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging cPILOT
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21-plex DiLeu Isobaric Tags for High-Throughput Quantitative Proteomics.

Dustin C Frost1, Yu Feng1, Lingjun Li1,2

  • 1School of Pharmacy, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.

Analytical Chemistry
|May 14, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed new DiLeu isobaric tags for cost-effective, 21-plex quantitative proteomics. These tags enable high-resolution mass spectrometry analysis of multiple samples in a single experiment, expanding multiplexing capacity.

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Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Isobaric tags are crucial for multiplexed quantitative proteomics.
  • Commercial reagents are often expensive.
  • Custom-developed DiLeu tags offer a cost-effective alternative.

Purpose of the Study:

  • To introduce a new generation of DiLeu isobaric tags.
  • To expand multiplexing capacity to 21 samples.
  • To maintain cost-effectiveness and performance.

Main Methods:

  • Synthesis of nine new reporter variants using stable isotopes (13C, 15N, 2H).
  • High-resolution HCD MS/MS analysis on Orbitrap platforms.
  • Single-shot nanoLC-MS/MS on a Q Exactive HF system.

Main Results:

  • Successful 21-plex quantification using distinct reporter ions with minimal mass differences (3 mDa).
  • Generation of strong reporter ions in HCD MS/MS spectra.
  • Accurate quantification of protein digests from K562 human cell line.

Conclusions:

  • The new 21-plex DiLeu tags significantly enhance multiplexing capacity for quantitative proteomics.
  • These tags provide a high-performance, low-cost solution for large-scale proteomic studies.
  • The developed method is suitable for accurate protein quantification in complex biological samples.