Quantitative Protein Analysis using SDS-PAGE

Analysis of protein pattern were performed as previously seen^{53 (link)}. Total protein concentrations in extracts were determined by the Bradford method with bovine serum albumin (BSA) as the standard. Then, SDS-PAGE gel electrophoresis was performed on samples with equalized concentrations of total protein. In Particular 20 μl of samples was mixed with 20 μl of Laemmli Sample Buffer with β-mercaptoethanol (Bio-Rad) and heating (95 °C, 5 min). then 40 μl of mixed samples and 5 μl of protein marker (Precision Plus Protein Dual Color Standards, Bio-Rad) were loaded onto the gel and resolved, using the Mini-PROTEAN electrophoresis system (Bio-Rad). The protein bands, separated on gel, were fixed, stained in QC Colloidal Coomassie Stain and destained solution according to the producer procedure (Bio-Rad).