Antibody Detection Assay for Adenovirus E1A

As performed previously [2 (link)], mouse monoclonal anti-E1A M58 antibody was previously described [13 (link)] and was grown in-house and used as the supernatant of M58 hybridoma cells. The M58 antibody binds within the region encoded by the first exon of E1A (between residues 63 and 99 of E1A) and recognizes all mutant E1A proteins used in this study [14 (link)]. Mouse monoclonal anti-72k DNA binding protein (DBP) antibody was previously described [15 (link)] and was used at a dilution of 1:400 for western blot. Anti-adenovirus type 5 antibody was purchased from Abcam (catalog number ab6982, Cambridge, MA, USA). Actin antibody was purchased from Abcam (catalog number ab3280). Secondary antibodies were acquired from Jackson ImmunoResearch (West Grove, PA, USA) and were used at a dilution of 1:200,000.