Western blot detection of recombinant hGAD65 from total soluble protein extracted from chloroplast transgenic algal lines. Samples containing ~50 ng of commercial hGAD65 or 5 μg of total algal protein were separated by 15% sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred onto a polyvinylidene difluoride membrane and probed with rabbit anti-GAD65 primary antibody followed by horseradish peroxidase conjugated goat anti-rabbit secondary antibody. Standard, insect cell-derived hGAD65. Lanes 1 to 2, protein extracts from two representative algal transformants. Protein extracts from wild-type strain 137c were used as a negative control. Arrows indicate the position of algal-derived hGAD65.
Wang et al. BMC Biotechnology 2008 8:87 doi:10.1186/1472-6750-8-87