A "de-streaking" method for two-dimensional electrophoresis using the reducing agent tris(2-carboxyethyl)-phosphine hydrochloride and alkylating agent vinylpyridine

Fengju Bai, Sheng Liu, Frank Witzmann

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Optimal isoelectric focusing in the alkaline region remains a challenge in two-dimensional gel electrophoresis (2-DE), though various attempts had been made to reduce basic end streaking. The present study reports the application of a novel reduction and alkylation step prior to 2-DE analysis using tris(2-carboxyethyl)-phosphine hydrochloride as a reducing agent and vinylpyridine as an alkylating agent. This simple sample preparation approach effectively eliminates basic end streaks, thereby enabling the analysis and identification of more protein spots resolved by 2-DE.

Original languageEnglish
Pages (from-to)2043-2047
Number of pages5
JournalProteomics
Volume5
Issue number8
DOIs
StatePublished - May 2005

Fingerprint

Alkylating Agents
Reducing Agents
Alkylation
Electrophoresis, Gel, Two-Dimensional
Isoelectric Focusing
Electrophoresis
Gels
Proteins
tris(2-carboxyethyl)phosphine

Keywords

  • De-streaking
  • Reduction and alkylation
  • Tris(2-carboxyethyl)-phosphine
  • Two-dimensional gel electrophoresis
  • Vinylpyridine

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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AU - Liu, Sheng

AU - Witzmann, Frank

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AB - Optimal isoelectric focusing in the alkaline region remains a challenge in two-dimensional gel electrophoresis (2-DE), though various attempts had been made to reduce basic end streaking. The present study reports the application of a novel reduction and alkylation step prior to 2-DE analysis using tris(2-carboxyethyl)-phosphine hydrochloride as a reducing agent and vinylpyridine as an alkylating agent. This simple sample preparation approach effectively eliminates basic end streaks, thereby enabling the analysis and identification of more protein spots resolved by 2-DE.

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