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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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Updated: Jul 11, 2025

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.

Melissa Petiti¹, Laetitia Houot¹, Denis Duché²

¹Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France.

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

|November 6, 2023

Summary

This summary is machine-generated.

This study presents a straightforward fractionation method to isolate cellular compartments in Escherichia coli. This technique aids in determining protein localization and distinguishing between integral and peripheral inner membrane proteins.

Keywords:

Freeze and thaw Membrane Osmotic shock Peptidoglycan Protein solubilization Spheroplast Subcellular localization

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Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

Published on: July 15, 2018

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Last Updated: Jul 11, 2025

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

Area of Science:

Microbiology
Molecular Biology
Biochemistry

Background:

Protein function is intrinsically linked to its subcellular location within a cell.
Gram-negative bacteria like Escherichia coli possess five distinct cellular compartments: cytoplasm, inner membrane, periplasm, outer membrane, and extracellular medium.

Purpose of the Study:

To describe a simple and efficient method for fractionating Escherichia coli cells.
To enable the determination of protein presence within specific cellular compartments.
To differentiate between integral and peripheral inner membrane proteins.

Main Methods:

Cellular fractionation of Escherichia coli.
Biochemical techniques for compartment isolation.
Analysis of protein presence in isolated fractions.

Main Results:

Successful isolation of distinct cellular compartments from Escherichia coli.
Demonstration of a method to identify protein localization.
A specific protocol to distinguish integral from peripheral inner membrane proteins.

Conclusions:

The described fractionation method is effective for analyzing protein localization in Escherichia coli.
This approach facilitates the study of protein targeting and function in gram-negative bacteria.
The method provides a means to characterize inner membrane protein types.