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Electrophoretic Separation of Proteins
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Sample preparation guidelines for two-dimensional electrophoresis.

Anton Posch1

  • 1Bio-Rad Laboratories GmbH , Heidemannstrasse Munich , Germany.

Archives of Physiology and Biochemistry
|September 12, 2014
PubMed
Summary
This summary is machine-generated.

This review offers essential guidelines for optimizing protein sample preparation for two-dimensional electrophoresis (2DE). It helps researchers avoid common pitfalls and develop effective protocols for gel-based proteomics and other protein separation techniques.

Keywords:
Cell disruptioncell lysis buffercontaminant removalstain-free technology

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Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Effective sample preparation is crucial for successful two-dimensional electrophoresis (2DE).
  • The diversity of protein samples necessitates empirical determination of optimal preparation methods.
  • Numerous potential pitfalls exist in protein sample preparation for proteomics.

Purpose of the Study:

  • To provide a comprehensive overview of key principles in protein sample preparation for 2DE.
  • To guide researchers in avoiding common errors during sample preparation.
  • To facilitate the development of straightforward protocols for gel-based proteomics.

Main Methods:

  • Screening and evaluation of expert-derived sample preparation protocols.
  • Consolidation of practical experience and established protocols.
  • Development of guidelines for rationalizing sample preparation choices.

Main Results:

  • Identification of critical principles for successful protein sample preparation.
  • Provision of practical guidelines to aid researchers in protocol development.
  • Strategies applicable to both 2DE and other protein separation techniques.

Conclusions:

  • Optimized sample preparation is fundamental for reliable 2DE results.
  • The guidelines presented simplify protocol development for researchers.
  • The reviewed strategies enhance protein sample preparation across various separation techniques.