Why does the PINK1 (D8G3) Rabbit mAb #6946 detect two bands for human PINK1?

In endogenous samples, the PINK1 precursor protein (~60kDa), is imported into mitochondria where it is processed into a cleaved form (~50kDa) that is transported back to the cytoplasm to be rapidly degraded by the proteasome. Since PINK1 is degraded so quickly, detecting endogenous PINK1 in untreated cell extracts is unlikely. In order to observe PINK1 in endogenous samples, we use the induction of mitophagy by treatment with CCCP, which leads to the accumulation of cleaved PINK1. 

However, we do observe two bands, representing both the precursor and cleaved forms, when using samples transfected with full-length PINK1. This is due to the high levels of PINK1 in these samples. 

Last updated: February 28, 2024