Figure 5
Ponceau S staining is comparable to β-actin as a loading control for immunobloting. HCT116 cells were plated at 106 per 100 mm plate, and cells were processed for protein extraction with standardized laboratory A+2X protocol, 48 h after plating. Increasing input of protein extract, from 20 to 140 μg with 20 μg increments, were next separated in 8% SDS-PAGE followed by transfer to nitrocellulose membrane, Ponceau S staining and β-actin steady-state expression evaluation. (A) Representative Ponceau S staining and steady-state expression levels of β-actin. (B) Correlation between Ponceau S and β-actin relative quantification, from three independent experiments.