Baculovirus Protein Expression Analysis

Baculoviruses were lysed with loading buffer containing 2% 2-mercaptoethanol, boiled for 5 min. The lysates were separated by 8% SDS-PAGE and transferred to polyvinylidene fluoride membranes, and then probed with either of the following: an anti-PfCSP mouse mAb (2A10) or an anti-hDAF mouse mAb (anti-CD55, Merck Millipore, Temecula, CA), together with an anti-VP39 rabbit Ab [14 (link)]. Blots probed with the appropriate secondary Abs conjugated to IRDye 680 and IRDye 800 (Rockland Immunochemicals, Gilbertsville, PA) in the same membranes were visualized using an Odyssey infrared imager (LI-COR, Lincoln, NE). The molecular weight predictions were carried out on the ExPASy server, and densitometry analyses were performed using Image Studio Digits (LI-COR).