Goat anti rabbit igg hrp secondary antibody

The protein levels of cleaved caspase-3, cleaved caspase-9, cleaved PARP, p21, p53, Bax, Bcl-2, HO-1, NQO1, Nrf2, AMPK, GSK-3β, p-AMPK, and p-GSK-3β in the SH-SY5Y cells were verified via western blot analysis. Equal quantities of protein samples resolved on 7.5–3% acrylamide sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels were transferred to polyvinylidene fluoride (PVDF) membranes. The antibody dilution ratios were as follows: α-tubulin (1:2,000), TBP (1:500), Nrf2 (1:500), NQO1 (1:500), and HO-1 (1:1,000; Santa Cruz Biotechnology, Inc.); cleaved caspase-3 (1:1,000), cleaved PARP (1:1,000), cleaved caspase-9 (1:1,000), p53 (1:1,000), p21 (1:1,000), Bax (1:1,000), AMPK (1:1,000), p-AMPK (1:1,000), GSK-3β (1:1,000), and p-GSK-3β (1:1,000; Cell Signaling Technology Inc.). Thereafter, the membranes were incubated with a 1:5,000 dilution of goat anti-rabbit IgG-HRP secondary antibody (Cell Signaling Technology Inc.). Data were scanned using an imaging system (Amersham Biosciences) and analyzed using the ImageQuant 350 analyzer.