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Comparative evaluation of extraction methods for total proteins from Crocus sativus L. (Saffron)
Authors:Syed Sana Mehraj  Azra N Kamili  Ruqeya Nazir  Ehtishamul Haq  Henah Mehraj Balkhi
Institution:1. Centre of Research for Development, University of Kashmir, Srinagar, J&K 190006, India;2. Department of Biotechnology, University of Kashmir, Srinagar, J&K 190006, India
Abstract:Broadly speaking proteomic studies are one of the various techniques of utmost importance for understanding complex biological processes that occur under inductive conditions and revealing the multidimensional aspects of Crocus sativus in biological systems. In order to get an insight into the molecular changes and to characterize the variations in protein expression of C. sativus, a detailed proteomic analysis on one-dimensional gel electrophoresis is one of the basic steps to accomplish. We have compared total protein profiles of C. sativus extracted by three different recipes and analyzed on 10% sodium dodecyl sulfate polyacrylamide gels. Gels were subjected to densitometric analysis for further characterization. Among three different protocols NP-40 extraction buffer recipe resulted in the extraction of proteins most efficiently with minimum background and streaking. There was maximum solubilization of proteins with high efficiency. Such a profile can be used for high precision analysis of differential protein expression. This work is an attempt to assist researchers in effective extraction of proteins from C. sativus. As a researcher faces a perplexing array of choices as where to start we describe a method based on our collective analysis of the different protein protocols. This paper presents a method that could be applied at the outset of any proteomic study.
Keywords:SDS–PAGE  sodium dodecyl sulfate polyacrylamide gel electrophoresis  1-DE  one dimensional gel electrophoresis  2-DE  two-dimensional gel electrophoresis  DTT  dithiothreitol  2-ME  2-mercaptoethanol  Proteomics  Proteins  Biological
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