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Subcellular Fractionation

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 Isolation

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: May 6, 2026

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

Published on: August 12, 2021

Cellular fractionation--yeast cells.

Joerg von Hagen¹, Uwe Michelsen

¹Merck KGaA, Darmstadt, Germany.

Methods in Enzymology

|November 5, 2013

Summary

This summary is machine-generated.

Subcellular fractionation is a key proteomics technique for isolating cellular components. This method reduces sample complexity, enhancing analysis of organelles and protein complexes.

Keywords:

Cytoskeleton extraction buffer Membrane protein extraction buffer Nuclear protein extraction buffer Protoplast buffer and cytoplasmic protein extraction buffer Subcellular fractionation of yeast protoplasts Yeast cells Yeast protoplast formation

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Area of Science:

Proteomics
Cell Biology
Biochemistry

Background:

Subcellular fractionation is a critical enrichment technique in proteomics.
Analyzing intracellular organelles and multiprotein complexes requires specialized methods.
Reducing sample complexity is essential for high-resolution proteomic analysis.

Purpose of the Study:

To highlight the importance of subcellular fractionation in proteomics.
To explain its role in analyzing specific cellular components.
To discuss its compatibility with various analytical techniques.

Main Methods:

Subcellular fractionation as an enrichment strategy.
Combination with high-resolution 2-D gel electrophoresis and mass spectrometry.
Integration with gel-independent proteomic techniques.

Main Results:

Subcellular fractionation effectively reduces sample complexity.
This technique is crucial for the detailed analysis of intracellular structures.
Optimized fractionation improves the efficiency of subsequent proteomic analyses.

Conclusions:

Subcellular fractionation is an indispensable technique in modern proteomics.
Its flexibility allows for adaptation to diverse research needs.
Combining fractionation with advanced analytical methods maximizes data quality and biological insight.