Imaging of Chemiluminescent Western Blots: Comparison of Digital Imaging and X-ray Film

The decay of signal over time was performed using serial dilution of a E. coli cell lysate containing a 51 kD polyhistidinetagged recombinant protein run on a 4–20% linear gradient Criterion gel. Blots were probed with mouse anti-polyhistidinetag primary antibody followed by incubation with GAM-HRP secondary antibody and StrepTactin HRP (1:10,000) for 1 hr each. The blots were then cut into two sections, each containing an identical dilution series of protein. The sections of membrane were incubated in chemiluminescent substrates.