Protein Extraction from Rose Cultivars

Protein extraction was done according to our previous methods [27 (link)]. The frozen powdered samples from all cut rose cultivars was extracted with extraction buffer (pH 7.5) containing 40 mM (w/v) Tris-HCl, pH 7.5, 2 mM (w/v) EDTA, 0.07% (w/v) β-mercaptoethanol, 2% (w/v) PVP and 1% (v/v) Triton X-100. The extract was centrifuged at 13,000 rpm for 10 min at 4 °C. The supernatant was mixed with protein-dye and 20 μg proteins were loaded on 12.5% polyacrylamide gel on PROTEAN II (Bio-Rad, Hercules, CA, USA). The protein concentration was determined by the Bradford method using BSA (bovine serum albumin) as a standard curve. After electrophoresis, the gels were stained with a commercial available silver stain according to manufacturer’s instructions (Bio-Rad, Hercules, CA, USA).