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A “de‐streaking” method for two‐dimensional electrophoresis using the reducing agent tris(2‐carboxyethyl)‐phosphine hydrochloride and alkylating agent vinylpyridine
Author(s) -
Bai Fengju,
Liu Sheng,
Witzmann Frank A.
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200501291
Subject(s) - streaking , reducing agent , tris , phosphine , electrophoresis , hydrochloride , isoelectric focusing , chemistry , chromatography , tcep , alkylation , combinatorial chemistry , organic chemistry , biochemistry , biology , microbiology and biotechnology , enzyme , catalysis
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.