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Published on: November 10, 2016
TCA precipitation and ethanol/HCl single-step purification evaluation: One-dimensional gel electrophoresis, bradford
Balkis Eddhif1, Nadia Guignard2, Yann Batonneau2
1Institut de Chimie des Milieux et des Matériaux de Poitiers (IC2MP), Université de Poitiers, CNRS, Equipe Eaux, Biomarqueurs, Contaminants Organiques, Milieux, (E-BiCOM), 4 rue Michel Brunet, TSA 51106, F-86073 Poitiers, France.
Abstract:
The data presented here are related to the research paper entitled "Study of a Novel Agent for TCA Precipitated Proteins Washing - Comprehensive Insights into the Role of Ethanol/HCl on Molten Globule State by Multi-Spectroscopic Analyses" (Eddhif et al., submitted for publication) [1]. The suitability of ethanol/HCl for the washing of TCA-precipitated proteins was first investigated on standard solution of HSA, cellulase, ribonuclease and lysozyme. Recoveries were assessed by one-dimensional gel electrophoresis, Bradford assays and UPLC-HRMS. The mechanistic that triggers protein conformational changes at each purification stage was then investigated by Raman spectroscopy and spectrofluorometry. Finally, the efficiency of the method was evaluated on three different complex samples (mouse liver, river biofilm, loamy soil surface). Proteins profiling was assessed by gel electrophoresis and by UPLC-HRMS.
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