Gfp d5.1 xp rabbit monoclonal antibody

Western blot analyses were performed as previously described (30 (link)). Polyclonal rabbit anti-FLAG antiserum (antibodies-online) was used at a dilution of 1:20 000 to detect RpsT-3xFLAG. GFP was detected using GFP (D5.1) XP^® rabbit monoclonal antibody (Cell Signaling Technology) diluted 1:10 000. The antibodies were visualized using goat anti-rabbit IgG secondary antibodies conjugated to alkaline phosphatase (Promega) at a dilution of 1:100 000. Signals were detected by chemiluminescence using CDP* as substrate (Novagen).

Japanese encephalitis virus GI strain SH7 (MH753129) and GIII strain SH15 (MH753130), isolated from Cx. tritaeniorhynchus and An. sinensis (respectively) in 2016 were used in this study [37 (link), 38 (link)]. All JEV strains were plaque-purified three times and amplified in BHK cells at 0.1 MOI as described previously [37 (link)]. The fifty percent tissue culture infective dose (TCID50) was determined, and fresh virus suspensions were used in all experiments. Baby hamster kidney cells (BHK-21) cell line was obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, GIBCO, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 100 μg/ml streptomycin and 100 IU/ml penicillin at 37°C. The commercial antibodies used in this study included a GFP (D5.1) XP Rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA), GAPDH (glyceraldehyde-3-phosphate dehydrogenase) monoclonal antibody (Proteintech, Chicago, IL, USA), monoclonal His-tag antibody (GeneScript, Piscataway, NJ, USA) and a self-generated antibody specific to JEV NS3 [39 (link)].