Figure 1. Specificity of PARP-1 (cleaved) antibody demonstrated by Western blot. Western blot analysis of HeLa cells either untreated (CON) or treated with 1 μM Staurosporine (STS) for 4 hours to induce apoptosis. This antibody recognizes cleaved PARP-1 (89 kDa) and not full length PARP-1 (113 kDa). Antibody was from MitoSciences PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215).
Western blots yield useful information about specificity, even though antigen presentation is different than in fixed cells.1,2
Figure 2. Inhibition of mitochondrial biogenesis by chloramphenicol. A) Western blot validation. Chloramphenicol treatment inhibits mtDNA-encoded COX-1 protein synthesis, but not nuclear-DNA-encoded SDH-A protein synthesis. B) The IC50 of the drug's effect on mitochondrial protein translation was determined with the Mitosciences MitoBiogenesis™ In-Cell ELISA Kit (IR) (ab110216). Cells were seeded at 3000 cells/well and allowed to grow for 3 cell doublings in a drug dilution series. Relative amounts of COX-1 and SDH-A were measured in each well. Chloramphenicol inhibits COX-1 protein synthesis relative to SDH-A protein synthesis by 50% at 3.5 μM.