A high protein recovery extraction method from buckwheat based on protein yield gel quality and 1-D gel electrophoresis

Proteomics is an OMICS based technology used to functional characterization of gene expression. Its potential depends on recovery of proteins from tissues. A protocol that will lead to maximum recovery of proteins from any tissue is considered as the best method and efficient means to proceed for in depth proteome analysis. In this respect, we have tried various methods to establish best protein extraction method for recovery of proteins from different tissues of buckwheat plant. The tissues that we considered are shoot, root and seed. In case of Buckwheat, this is the first report in which we have established protein extraction methods from various tissues. Extracted proteins were furthered quantified using Bradford method and then resolved on SDS-PAGE. The results indicated that the efficacy of a specific protein extraction methodology is dependent on the structure and metabolic contents of different tissues. Among various methods tested phosphate -TCA acetone method showed high recovery of proteins from buckwheat shoots and roots having highest protein concentration of 3.31±0.05μg/μl and 1.20±0.04μg/μl in shoots and roots respectively. Whereas, by phenol-based protocol protein yield of 3.61±0.07 μg/μl was obtained from buckwheat seeds.