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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
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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: Jun 3, 2026

Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana

Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana

Published on: August 17, 2009

Plant protoplast enucleation by density gradient centrifugation.

H K Fakhrai¹, N Haq, P K Evans

¹University of Durham, Durham, UK.

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

|March 11, 2011

Summary

This summary is machine-generated.

Cell enucleation can be achieved by centrifuging cells in density gradients, separating nuclei from cytoplasm. This method is less explored in plant cells compared to mammalian cells.

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Area of Science:

Cell biology
Biotechnology

Background:

Cell enucleation methods are crucial for various biological studies.
Mammalian cell enucleation has been achieved through surface adhesion or density gradient centrifugation.

Purpose of the Study:

To explore density gradient centrifugation as a method for cell enucleation.
To investigate the generation of cytoplasts and mini-cells.

Main Methods:

Cells are centrifuged at high speed within a density gradient.
Differential densities of nucleus and cytoplasm are exploited for separation.
Stretching forces generated during centrifugation facilitate enucleation.

Main Results:

Density gradient centrifugation effectively separates nuclei from cytoplasm.
High percentages of enucleated cells (cytoplasts) and mini-cells can be produced.
This technique is less established in plant cells than in mammalian cells.

Conclusions:

Density gradient centrifugation is a viable method for cell enucleation.
Further research is warranted to explore its application in plant cells.