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Updated: Jun 19, 2026
09:20
In situ Subcellular Fractionation of Adherent and Non-adherent Mammalian Cells
Published on: July 23, 2010
Isolation of subcellular organelles and structures
Uwe Michelsen1, Jörg von Hagen
1Merck KGaA, Darmstadt, Germany.
Methods in Enzymology
|November 7, 2009
Summary
This study details methods for isolating specific cellular components in proteomics. These techniques improve the detection and analysis of low-abundance proteins in complex biological samples.
Area of Science:
- Proteomics
- Cell Biology
- Biochemistry
Background:
- Functional proteomics requires separating complex protein mixtures for accurate analysis.
- Standardized sample preparation is crucial for reproducible protein separation and detection.
- Analyzing total proteomes is challenging due to complexity and protein property variations.
Purpose of the Study:
- To describe detailed procedures for enriching and isolating organelles and subcellular compartments.
- To enhance the coverage and detection of low-abundance proteins in mammalian cells.
- To provide standardized methods for functional proteomic analyses.
Main Methods:
- Classical centrifugation techniques for organelle isolation.
- Sophisticated immunoaffinity-based procedures for subcellular compartment enrichment.
- Detailed recipes for sample preparation prior to protein separation.
Main Results:
- Established protocols for isolating key mammalian cell organelles and subcellular compartments.
- Demonstrated improved visualization and accessibility of low-abundance proteins.
- Provided standardized procedures for reproducible proteomic analyses.
Conclusions:
- Enrichment of specific proteomes is beneficial for comprehensive proteomic studies.
- Standardized isolation techniques are essential for reliable functional proteomics.
- The described methods facilitate advanced analysis of proteins affected by environmental factors.
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