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Updated: Jul 2, 2026
Glycopeptide Capture for Cell Surface Proteomics
Published on: May 9, 2014
Sample preparation method for plasma membrane proteome analysis
Seung-Ah Park1, Mi-Ryung Kim, Pan-Kyeom Kim
1School of Life Sciences and Biotechnology, Korea University, 1, 5-ga Anam-dong, Sungbuk-ku, Seoul 136-701, Republic of Korea.
Abstract:
The preparation of plasma membrane (PM) proteome samples is seriously difficult and time-consuming, owing to their profound hydrophobicity and low abundance. We have developed an efficient PM sample preparation method using Ultracentrifugation with Percoll and an aqueous two-phase extraction. The developed method was rapid (3 h) and provided high purities (26-fold of cell lysate) with a high yield (2.6% of whole cell lysate proteins). This method is especially useful for PM proteome studies using 2D gel electrophoresis.
Insights
Preparing plasma membrane (PM) proteome samples is challenging. A new method using Ultracentrifugation with Percoll and aqueous two-phase extraction offers a rapid, high-purity, and high-yield solution for PM proteome studies.
Area of Science:
- Proteomics
- Cell Biology
- Biochemistry
Background:
- Plasma membrane (PM) protein isolation is crucial for cell biology research.
- PM proteins are hydrophobic and low in abundance, making sample preparation difficult and time-consuming.
Purpose of the Study:
- To develop an efficient, rapid, and high-purity method for plasma membrane proteome sample preparation.
- To facilitate downstream analyses such as 2D gel electrophoresis.
Main Methods:
- Ultracentrifugation with Percoll combined with aqueous two-phase extraction.
- Optimization of sample preparation for plasma membrane proteins.
Main Results:
- Achieved a rapid sample preparation time of 3 hours.
- Obtained high purity, with a 26-fold enrichment compared to cell lysate.
- Secured a high yield, representing 2.6% of whole cell lysate proteins.
Conclusions:
- The developed method provides an efficient and effective approach for plasma membrane proteome analysis.
- This technique is particularly beneficial for studies utilizing 2D gel electrophoresis.
- The method addresses the challenges of hydrophobicity and low abundance in PM protein isolation.
