Anti mev nucleocapsid

Cells grown on a 6-well plate were infected with various rMeVs at an MOI of 0.03. At 36–48 h post-infection, the supernatant was collected and filtered through a 0.45-μm pore membrane. Additionally, the cells were lysed in mammalian protein extraction reagent (Cat# 78503, Thermo Fisher Scientific) supplemented with Halt protease inhibitor cocktail (Cat# 877886, Thermo Fisher Scientific). The protein concentration was determined using a Pierce Coomassie Plus Assay Kit (Cat# 23236, Thermo Fisher), and 3 µg of cell lysate or ~20 µL of supernatant was separated on a precast 12% or 4%–12% Bis-Tris polyacrylamide gel before being transferred to a polyvinylidene fluoride membrane using an iBlot2 dry blotting system (Thermo Fisher Scientific). The blot was then probed with anti-SARS-CoV-2 spike RBD (GTX135385, GeneTex, Irvine, CA, USA), anti-SARS-CoV-2 spike (Cat# GTX632604, GeneTex), anti-MeV nucleocapsid (Cat# LS-C144599, LsBio, Seattle, WA, USA), anti-MeV hemagglutinin (116 (link)), anti-canine distemper virus fusion (Genscript), and anti-high affinity (HA) peroxidase (Cat#12013819001, Millipore Sigma, St. Louis, MO, USA) and developed with a KwikQuant Western Blot Detection Kit using a KwikQuant Imager (Kindle Bioscience LLC, Greenwich CT, USA).