Irdye fluor 800 labeled igg secondary antibody

Western blot assay was carried out as follow: cells were lysed with RIPA buffer (Santa Cruz, USA) and cleared lysate was collected by centrifugation for protein separation on 10% SDS-polyacrylamide gel. The proteins were transferred onto PVDF membranes (Millipore) and detected with respective antibodies at 4°C overnight, followed by incubation with either IRDye Fluor 680-labeled IgG or IRDye Fluor 800-labeled IgG secondary antibody (Li-Cor Bioscience). The images were scanned and quantified by densitometric analysis by Li-COR Odyssey Infrared Imager. Primary antibodies against VP1 (Millipore), CD81 (Cell Signaling Technology), IRAK1 (Santa Cruz), CD63 (Abcam), ISG15 (Abcam) and Rab27a, TRAF6, STAT1, TSG101, BST-2/Tetherin, and GAPDH (all from Proteintech), VP2, 3AB, 3C and 3D (all from Genetex) were used.

The information of anti-HSV-1 ICP0, anti-HSV-1 ICP4, anti-HSV-1/2 gD, and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) rabbit is shown in Table 3. DAPI (4′,6-diamidino-2-phenylindole) (Thermo Fisher Scientific) was used as a counter to stain the nuclei, IRDye Fluor 680-labeled IgG or IRDye Fluor 800-labeled IgG secondary antibody (LI-COR Bioscience).