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Subcellular Fractionation01:32

Subcellular Fractionation

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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...
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Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

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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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Centrifugation01:05

Centrifugation

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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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Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
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Related Experiment Video

Updated: Oct 11, 2025

Phase Behavior of Charged Vesicles Under Symmetric and Asymmetric Solution Conditions Monitored with Fluorescence Microscopy
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Phase Behavior of Charged Vesicles Under Symmetric and Asymmetric Solution Conditions Monitored with Fluorescence Microscopy

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Current methods for studying intracellular liquid-liquid phase separation.

Amber R Titus1, Edgar E Kooijman1

  • 1Department of Biological Sciences, Kent State University, Kent, OH, United States.

Current Topics in Membranes
|December 4, 2021
PubMed
Summary
This summary is machine-generated.

Liquid-liquid phase separation (LLPS) forms membrane-less compartments crucial for cell signaling. Understanding LLPS mechanisms and modeling these intracellular phase transitions are key to deciphering cellular processes and dysregulation.

Keywords:
Aqueous two-phase systemsInterfacial tensionIntrinsically disordered proteinLiquid-liquid phase separationMembrane-less organelle

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Biophysics

Background:

  • Liquid-liquid phase separation (LLPS) drives the formation of membrane-less intracellular compartments.
  • These compartments concentrate biomolecules, influencing cellular signaling.
  • Dysregulation of LLPS is implicated in various diseases.

Purpose of the Study:

  • To highlight recent advances in understanding intracellular phase transitions.
  • To review current methods for identifying LLPS in vitro.
  • To discuss models for studying LLPS in situ.

Main Methods:

  • Review of recent literature on LLPS.
  • Description of experimental techniques for in vitro LLPS identification.
  • Overview of computational and imaging approaches for in situ LLPS modeling.

Main Results:

  • LLPS is a fundamental mechanism for cellular organization.
  • Diverse methods exist to detect and characterize phase-separated biomolecular condensates.
  • In situ modeling provides insights into dynamic LLPS processes within cells.

Conclusions:

  • LLPS is critical for cellular function and signaling.
  • Advanced techniques are available for studying LLPS.
  • Further research into LLPS mechanisms and modeling is essential for understanding cellular regulation and disease.