The densitometry and the concentration of proteins in seeds dehydrated by TSD and DEMI-LP were determined.
With the protein extracts obtained, the 12% PAGE-SDS polyacrylamide gels were made (Acrylamide-Bis Acrylamide 30:08, tris–HCl 1.5 M pH 8.8, SDS 10%) by conventional methods, and 60 mg of protein per lane was used. These samples were denatured for 5 min with a denaturing buffer (2.5 mL Tris–HCl 0.5 M pH 6.8,1 mL Bis-mercaptoethanol 5%, 4.5 mL distilled H2O, 4 mL 10% SDS, 20 μL or 0.0002 g bromophenol blue 1%, 8 mL Glycerol 10%) consisting in tubes placed over boiling water in a hot dish. The gels were run for 30 min at 80 V and then for 120 min at 100 V. Once the shift was over, the gels were stained with Coomassie blue G-250 (C47 H48 N3 O7 S2 Na ) in an aqueous acetic acid solution (10%).
With the protein extracts obtained, the 12% PAGE-SDS polyacrylamide gels were made (Acrylamide-Bis Acrylamide 30:08, tris–HCl 1.5 M pH 8.8, SDS 10%) by conventional methods, and 60 mg of protein per lane was used. These samples were denatured for 5 min with a denaturing buffer (2.5 mL Tris–HCl 0.5 M pH 6.8,1 mL Bis-mercaptoethanol 5%, 4.5 mL distilled H2O, 4 mL 10% SDS, 20 μL or 0.0002 g bromophenol blue 1%, 8 mL Glycerol 10%) consisting in tubes placed over boiling water in a hot dish. The gels were run for 30 min at 80 V and then for 120 min at 100 V. Once the shift was over, the gels were stained with Coomassie blue G-250 (C47 H48 N3 O7 S2 Na ) in an aqueous acetic acid solution (10%).
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