Fluorescein isothiocyanate fitc conjugated goat anti mouse igg antibody

HEK293T cells were transiently cotransfected with pCAGGS-E2-Flag (1 μg) and pMyc-MEK2 (1 μg). PK-15 cells were mock infected or infected with CSFV at an MOI of 0.1. At 36 hpt or 48 hpi, the plasmid-transfected or CSFV-infected cells were fixed with 4% paraformaldehyde and permeabilized with 0.15% Triton X-100. The cells were further incubated with a mouse anti-Flag MAb (catalog no. F1804; Sigma-Aldrich) or anti-E2 MAb HQ06 (31 (link)) for 1.5 h, followed by incubation with a rabbit anti-Myc PAb (catalog no. C3956; Sigma-Aldrich) or a rabbit anti-MEK2 PAb (catalog no. sc-525; Santa Cruz). Subsequently, the cells were incubated with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody (catalog no. F2012; Sigma-Aldrich) and tetramethyl rhodamine isocyanate-conjugated goat anti-rabbit IgG antibody (whole molecule) (catalog no. T6778; Sigma-Aldrich). After incubation with 4,6-diamidino-2-phenylindole (DAPI), the cells were observed using a Leica SP2 confocal system (Leica Microsystems; Germany).

BHK-21 and BSR/T7 cells [15 (link)] were propagated as described previously [16 (link)]. BSR/T7 cells were used to recover the recombinant virus and BHK-21 cells were used for titration, in vitro growth, and plaque assays.
WT FMDV r-HN (referred to here only), derived from a plasmid encoding the complete FMDV O/HN/CHA/93 genome (pOFS) [14 (link)], was passaged four times in BHK cells to further experiments.
MAb 3A24 directed against AEKNPLE (residues 99–105) epitope in NSP 3A of FMDV and MAb 3B4B1 directed against GPYAGPMER (residues 1–9) epitope in NSP 3B2 were obtained from the Lanzhou Veterinary Research Institute (LVRI). The region “AEKNPLE” was proven to be an immunodominant B-epitope in 3A protein by detecting the ability of FMDV reference sera recognized this region using a peptide ELISA in our lab (data not yet published). Fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody was purchased from Sigma.