Triton X-114 cloud point extraction to subfractionate blood plasma proteins for two-dimensional gel electrophoresis

2015 ◽  
Vol 485 ◽  
pp. 11-17 ◽  
Author(s):  
Flemming Jessen ◽  
Tune Wulff
Keyword(s):  
Blood Plasma ◽  
Cloud Point ◽  
Gel Electrophoresis ◽  
Plasma Proteins ◽  
Cloud Point Extraction ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Blood Plasma Proteins ◽  
Triton X

Effect of Triton X-100 and Dtt Concentrations on Wide Range Two-Dimensional Gel Electrophoresis of Tissue, Cell and Fluid Proteomes

2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Sébastien Charneau ◽  
Gabriel Costa Nunes da Cruz ◽  
Camila Miranda Costa ◽  
Marcelo Valle de Sousa ◽  
Carlos André Ornelas
Keyword(s):  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Water Soluble ◽  
Tissue Cell ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Ph Gradients ◽  
Tissue Samples ◽  
Wide Range ◽  
Triton X

AbstractHigh-resolution separation by two-dimensional gel electrophoresis (2- DE) is still challenging due to the intrinsic behavior of proteins, principally throughout isoelectric focusing separation. It is often observed low resolution of proteins in the alkaline pH region when using wide range pH gradients. Herein, we show the effect of different concentrations of Triton X‑100 and DTT in the sample buffer on wide range pH (3-10) 2-DE profiles of three different biological samples as Trypanosoma cruzi cells, honey bee brain tissue and human saliva fluid. Higher resolution, number and intensity of spots were achieved when 85 mM DTT and 2.5% Triton X‑100 were employed for cell and tissue samples. No improvement was observed for fluid proteins, probably because water-soluble proteins do not require special conditions for extraction and prevention of precipitation during isoelectric focusing.

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