A high protein recovery extraction method from buckwheat based on protein yield gel quality and 1-D gel electrophoresis Hami Ammarah1, Manzoor Madhiya1, Sudan Jebi1, Batool Aneesa1, Bashir Saika1, Zaffar Aaqif1, Sofi Parvaze Ahmad2, Afroza Baseerat3, Masoodi Khalid Z1, Bhat Sajad Ahmad4, Murtaza Imtiyaz4, Bhat M. Ashraf1, Khan Imran5, Zargar Sajad Majeed1,* 1Proteomics Laboratory, Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190 025, Jammu and Kashmir (India) 2Division of Genetics and Plant Breeding, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190 025, Jammu and Kashmir (India) 3Division of Vegetable Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190 025, Jammu and Kashmir (India) 4Division of Basic Science, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190 025, Jammu and Kashmir (India) 5Division of Statistics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190 025, Jammu and Kashmir (India) *e-mail: smzargar@skuastkashmir.ac.in
Online Published on 28 December, 2023. Abstract 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. Top Keywords 1D- PAGE, Buckwheat, Protein extraction methods, Protein quantification. Top |