Detection of EBV Latent Proteins by Western Blotting

Western blotting is a well established technique for identifying Epstein-Barr virus (EBV)-encoded proteins in lysates from cell lines or biopsy material. The basic technique involves separation of proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transfer onto a support membrane such as nitrocellulose or polyvinylidine difluoride (PVDF), followed by immunostaining with specific antibody reagents. The initial SDS-PAGE separation procedure is essentially similar to the method originally described by Laemmli (1 ), except that vertical slab gels are used instead of the original tube gels. Until recently, the slab gels commonly used were relatively large (approx 16�16 cm), but now it has become popular to use “mini” slab gels (approx 7�8 cm) because the electrophoresis and blotting times are considerably reduced and there is a significant saving in the amounts of the reagents required.