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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.

Centrifugation

Centrifugation

Centrifugation is a separation technique based on differences in density or size. It is commonly used to separate solids from aqueous interferents. During centrifugation, the sample is placed in centrifugation tubes and spun at high angular velocity, which allows centrifugal force to act differentially on the different densities or masses of the components. After spinning, the supernatant liquid is decanted. Depending on the specific application, either the pellet or the supernatant is retained...

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Related Experiment Video

Updated: Apr 30, 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

Cell fractionation.

Bérengère Ize¹, Véronique Viarre, Romé Voulhoux

¹Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255 - CNRS - Aix Marseille Université, IMM 31, Ch. J. Aiguier, 13402, Marseille, CEDEX 20, France, bize@imm.cnrs.fr.

Methods in Molecular Biology (Clifton, N.J.)

|May 14, 2014

Summary

This study details methods for isolating cellular compartments in Pseudomonas aeruginosa. This protein fractionation is key to understanding protein function within bacterial cells.

Area of Science:

Microbiology and Cell Biology
Bacterial Protein Localization

Background:

Proteins are localized to specific cellular compartments for distinct functions.
Gram-negative bacteria like Pseudomonas aeruginosa possess five distinct protein localization sites: cytoplasm, inner membrane, periplasm, outer membrane, and extracellular environment.

Purpose of the Study:

To describe methods for selectively isolating subcellular compartments in Pseudomonas aeruginosa.
To enable accurate determination of protein localization for functional studies.

Main Methods:

Cellular disruption techniques.
Centrifugation for selective isolation of subcellular compartments.
Protein fractionation for enrichment.

Main Results:

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Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

Published on: July 15, 2018

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Last Updated: Apr 30, 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

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

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

Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

Published on: July 15, 2018

Isolation of five distinct cellular compartments (cytoplasm, inner membrane, periplasm, outer membrane, extracellular environment) in Pseudomonas aeruginosa.
Enrichment of proteins within their respective subcellular compartments.

Conclusions:

Subcellular fractionation is essential for accurate protein localization studies in bacteria.
Understanding protein localization is a critical first step in elucidating protein function within Pseudomonas aeruginosa.