Thermo scientific pierce bicinchoninic acid bca protein assay kit

Total cellular protein was extracted using Pierce RIPA buffer (Thermo Fisher Scientific) supplemented with 1X cOmplet Mini Protease Inhibitor Cocktail (Roche) and 1X PhosSTO Phosphatase Inhibitor Cocktail (Roche). Protein quantification was performed using a Thermo Scientific Pierce Bicinchoninic Acid (BCA) protein assay kit (Thermo Fisher Scientific) according to manufacturer’s instructions and the absorbance was measured with an iMark microplate reader (Bio-Rad Laboratories, Hercules, CA).

The total RNA content for each sample of VLPs was determined by extracting the RNA from particle lysates with RNA columns, using Roche’s High Pure Viral RNA Kit (Roche Diagnostics, Indianapolis, IN, USA). The extracted RNA was detected by UV absorption at 260 nm using a Beckman DU-65 spectrophotometer (Beckman Coulter, Brea, CA, USA). A Thermo Scientific Pierce bicinchoninic acid (BCA) Protein Assay Kit (Thermo Fisher Scientific) was used to estimate the protein content of the same sample used to determine VLP RNA content. A bovine serum albumin protein standard and measurement of UV absorption at 562 nm were used to establish an approximate absorbance-to-concentration curve and estimate the total protein concentration of VLP samples.