A "de-streaking" method for two-dimensional electrophoresis using the reducing agent tris(2-carboxyethyl)-phosphine hydrochloride and alkylating agent vinylpyridine.
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.
Bai
Fengju
F
Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, 46202, USA.
Liu
Sheng
S
Witzmann
Frank A
FA
D000477:Alkylating Agents / Q000737:chemistry*; D000478:Alkylation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010720:Phosphines; D011725:Pyridines; D019163:Reducing Agents / Q000737:chemistry*
D000477:Alkylating Agents; D010720:Phosphines; D011725:Pyridines; D019163:Reducing Agents