Anti plk 1 rabbit polyclonal antibody

After 3 days of treatment under different conditions (pure media and media including UCNP@PEI + NIR, CRISPR-Cas9@PEI + NIR, UCNPs-Cas9@PEI, and UCNPs-Cas9@PEI + NIR), cells in six-well plates (5 × 10⁵ cells per well, before treatment) were washed with PBS twice and lysed on ice for 15 min with gentle stirring. After centrifugation at 12,000 rpm, a BCA protein assay kit (Sangon Biotech Co. Ltd., Shanghai, China) was applied to measure the total protein concentration of the supernatant cell lysates. Then, equal amounts of total protein (50 μg) were analyzed on 8% SDS–polyacrylamide gel electrophoresis and transferred onto a polyvinylidene difluoride membrane. The samples were blocked with the QuickBlock Blocking Buffer (Beyotime, China) for 1 hour at room temperature. After that, the membranes were incubated with 1000× dilutions of anti–glyceraldehyde-3-phosphate dehydrogenase and anti–PLK-1 rabbit polyclonal antibody (Sangon Biotech Co. Ltd., Shanghai, China), respectively, overnight, and immersed with the conjugated secondary antibody (Sangon Biotech Co. Ltd., Shanghai, China) at room temperature for 1 hour. Last, the membranes were washed five times in Tris-Buffered Saline with 0.02% Tween 20 buffer (5 min each) and targeted proteins were visualized by W-TMB (Sangon Biotech Co. Ltd., Shanghai, China).