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Related Concept Videos

Subcellular Fractionation01:32

Subcellular Fractionation

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...
Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

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Related Experiment Video

Updated: Jul 18, 2026

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification
10:19

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

Published on: August 12, 2021

Subcellular fractionation.

Mariana Eça Guimarães de Araújo1, Lukas Alfons Huber

  • 1Insbruck Biocentre, Austria.

Methods in Molecular Biology (Clifton, N.J.)
|December 19, 2006
PubMed
Summary

Highly sensitive mass spectrometry combined with subcellular fractionation enables detection of low-abundance regulatory proteins in organelles. This chapter details advanced protocols for protein analysis in cellular and tissue contexts.

Area of Science:

  • Proteomics
  • Cell Biology
  • Biochemistry

Background:

  • Detecting dynamic changes in low-abundance regulatory proteins within organelles is crucial for understanding cellular processes.
  • Subcellular fractionation is a widely adopted enrichment method due to its flexibility and ability to maintain protein context.
  • Mass spectrometry offers high sensitivity for protein detection.

Purpose of the Study:

  • To introduce state-of-the-art subcellular fractionation protocols.
  • To discuss the suitability, advantages, and limitations of various fractionation techniques.
  • To highlight the combined power of mass spectrometry and fractionation for organelle-level protein analysis.

Main Methods:

  • Subcellular fractionation of cells and tissues.

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Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation
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Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

Published on: January 18, 2019

Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens
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Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

Published on: July 15, 2018

Related Experiment Videos

Last Updated: Jul 18, 2026

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification
10:19

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

Published on: August 12, 2021

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation
06:53

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

Published on: January 18, 2019

Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens
10:07

Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

Published on: July 15, 2018

  • Pre-fractionation techniques for sample enrichment.
  • High-sensitivity mass spectrometry for protein detection and quantification.

    • Main Results:

    • Demonstration of successful combination of mass spectrometry and pre-fractionation.
    • Detection of dynamic changes in low-abundance regulatory proteins.
    • Analysis of proteins within their physiological/intracellular context at the organelle level.

    Conclusions:

    • Subcellular fractionation is a key preparative method for organelle proteomics.
    • Advanced protocols enhance the detection of low-abundance regulatory proteins.
    • The integration of fractionation and mass spectrometry provides powerful insights into cellular regulation.