Western Blot analysis of Mouse V5 tag - FITC Mouse Monoclonal Antibody
Western blot analysis of V5 tag was performed by loading 20 µg of whole cell extracts of untransfected HEK-293 and HEK-293 transiently overexpressing V5-His- LacZ using Novex® NuPAGE® 4-12% Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), and Novex® Sharp Pre-Stained Protein Standard (LC5800). Proteins were transferred to a PVDF membrane and blocked with 5% skim milk for 1 hour at room temperature. V5-His- LacZ was detected at ~117 kDa using Mouse V5 tag - FITC Mouse Monoclonal Antibody (R96325) at 1:3000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. Detection was performed and image captured at an FITC emission wavelength of 518nm.