Anti hsv 1 serum

All samples were lysed in SDS–PAGE loading buffer, sonicated, centrifuged, and denatured with 0.1 mol/L dl-dithiothreitol (Sigma-Aldrich) before loading. SDS-PAGE was conducted on 8%–18% gradient gels (GE Amersham Pharmacia Biotech, Munich, Germany), and proteins were electrotransferred onto nitrocellulose membranes (Bio-Rad, Hercules, CA). Membranes were incubated with the anti-HSV-1 serum (1:300, Dako) overnight at 4 °C. After washing, the blots were incubated with horseradish peroxidase-coupled sheep anti-rabbit secondary antibody (Amersham, Little Chalfont, UK) for 1 hour at room temperature, and reaction products were detected by chemiluminescence with the ImmunStar Western C Substrate kit (Bio-Rad) according to the manufacturer’s instructions. Ponceau S staining was included as loading control.