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Related Concept Videos

Proteomics01:33

Proteomics

7.3K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
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Subcellular Fractionation01:32

Subcellular Fractionation

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The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...
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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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Related Experiment Video

Updated: Jun 30, 2025

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier
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Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier

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Single Cell Analysis of Proteoforms.

Pei Su1, Michael A R Hollas1, Fatma Ayaloglu Butun2

  • 1Departments of Molecular Biosciences, Chemistry, Chemical and Biological Engineering, and Feinberg School of Medicine, Northwestern University, Evanston, Illinois 60208, United States.

Journal of Proteome Research
|March 18, 2024
PubMed
Summary
This summary is machine-generated.

We developed single cell Proteoform imaging Mass Spectrometry (scPiMS) for scalable single cell proteomics. This method enhances throughput and protein coverage by directly analyzing intact proteins from single cells.

Keywords:
individual ion mass spectrometrymass spectrometrynano-DESIproteoform assignment scoresingle celltop-down proteomics

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Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Cell Biology

Background:

  • Single cell proteomics is crucial for understanding cellular heterogeneity.
  • Current methods face throughput limitations, hindering large-scale analysis.

Purpose of the Study:

  • To introduce a novel platform for high-throughput single cell proteomics.
  • To enable direct analysis of intact proteins and their proteoforms within single cells.

Main Methods:

  • Developed single cell Proteoform imaging Mass Spectrometry (scPiMS).
  • Utilized direct solvent extraction and MS detection of intact proteins from single cells.
  • Employed drop-casting of single cells onto glass slides for sample preparation.

Main Results:

  • Achieved scalable single cell proteomics through intact proteoform analysis.
  • Significantly boosted cell processing rates by several fold.
  • Increased protein composition coverage compared to existing methods.

Conclusions:

  • scPiMS addresses the throughput bottleneck in single cell proteomics.
  • The platform enables efficient and comprehensive proteoform analysis at the single cell level.
  • scPiMS offers a scalable solution for advancing single cell proteomics research.