Figure 5
Protein quantity and quality from different melanin-bleaching methodologies. (a) Quantity of protein extracted under each condition was measured using a BCA Protein Assay kit. The protein extraction yield is expressed as the mean of three replicates (mean±s.d.). Black bars and clear bars represent the result of non-bleaching (▪) and bleaching (□) method, respectively. (b) Protein integrity under different melanin-bleaching conditions was assessed by gel electrophoresis. Proteins extracted after the use of different bleaching solutions were separated by 4–12% reducing SDS-PAGE and the gel was stained using a silver staining kit. (c) Proteins (10 μg) were subjected to 4–12% gradient SDS-PAGE, transferred to nitrocellulose membrane, and subjected to western blotting with anti-AKT (▪) and anti-GAPDH (□) antibodies. The signal was detected with a SuperSignal Chemiluminescence kit. NB, non-bleaching; Mr, protein molecular marker (kDa).
