Title:A Combined Method of Protein Extraction from Unorthodox Plant Samples for Proteomics
Volume: 18
Issue: 5
Author(s): Can Yilmaz*Mesude Iscan
Affiliation:
- Department of Molecular Biology and Genetics, Faculty of Science, Yuzuncu Yil University, Van,Turkey
Keywords:
2-DE, IEF, pinus, protein chip, proteomics, SDS, protein extraction.
Abstract:
Aims: This study aimed to generate an improved method of protein extraction and purification
from plant tissues containing very high amounts of phenolic compounds and other interfering
biomolecules.
Background: Protein extraction at proteomic studies on some plant species, including conifers, is
challenging, and the yield and quality are unpredictable.
Objective: Two popular protocols were combined with each other to construct a novel one with enhanced
abilities to produce higher purity of samples compatible for high precision molecular systems
and analysis.
Methods: The new method was compared with the other two for their efficiencies in classical SDS-
PAGE, 2-DE and capillary chromatography applications.
Results: All three methods were comparable in SDS-PAGE procedure; however, only the new
method created acceptable gel images in 2-DE. Bioanalyzer results, also, demonstrated that the
new method provided protein samples pure enough to be used in capillary chromatography with 2
times more peaks in electropherograms with lower noise and higher total relative protein concentrations
closest to the applied amount.
Conclusion: The new combined method is a successful alternative for plant proteomicists with
higher yield and quality of proteins from recalcitrant tissues.
Other: The new method could be preferred, especially, for high-tech, sensitive proteomic analysis.
