摘要
1.4 Western Blot For the analysis of mitochondrial proteins,organelles were first isolated from CD8^(+)T cells using the[Cell Mitochondria Isolation Kit,C3601,Beyotime,Shanghai,China]according to the manufacturer's instructions.Western Blot analysis was then performed on lysates from these purified mitochondrial fractions.Total protein was extracted from the cells mitochondria,added to 5×SDS loading buffer,and boiled at 100°C for 10 minutes.The separation glue was prepared as needed,ethanol was added,and the mixture was allowed to stand for 20 minutes.The separation glue was allowed to solidify,the ethanol was removed,the remaining ethanol was rinsed with water,and the residual liquid was removed with absorbent paper.
