Alexa fluor 488 conjugated goat anti mouse igg antibody

Mouse hybridomas producing the monoclonal antibody 4G2 anti-Flavivirus E were purchased from ATCC and a highly purified antibody preparation was produced by RD Biotech. The anti-mosquito saliva antibody was produced in house in rabbits exposed to mosquito bites. Horseradish peroxidase (HRP)-conjugated goat anti-mouse and anti-rabbit IgG antibodies were obtained from Bio-Rad Laboratories. Alexa Fluor 488-conjugated goat anti-mouse IgG antibody was obtained from Jackson ImmunoResearch.

Mosquito Aedes albopictus C6/36 cells were maintained at 28°C in Leibovitz medium (L15) supplemented with 10% heat-inactivated fetal bovine serum (FBS). Vero cells and were maintained at 37°C in Dulbecco’s modified Eagle medium (DMEM) supplemented with 5% FBS. BHK-21, SK-N-SH, and HEK-293T cells were maintained at 37°C in DMEM supplemented with 10% FBS. Highly purified mouse anti-pan Flavivirus E monoclonal antibody (mAb) 4G2 was produced by RD Biotech (Besançon, France). Mouse mAb anti-JEV NS5 was kindly provided by Y. Matsuura [26 (link)]. Antibodies against Calnexin and SNAP-tag were purchased from Enzo Life Sciences and New England Biolabs, respectively. Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG and anti-rabbit IgG antibodies were obtained from Bio-Rad Laboratories. HRP-conjugated goat anti-pig antibody was obtained from Bethyl Laboratories. Alexa Fluor 488-conjugated goat anti-mouse IgG antibody was obtained from Jackson ImmunoResearch.

HEK-293T cells from American Type Culture Collection (ATCC; Manassas, VA, USA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St Louis, MO, USA) and 1% penicillin-streptomycin (Thermo Fisher Scientific Waltham, MA). Coelenterazine h (Maokang Biotechnology, Shanghai, China) was used for the gaussia luciferase assay. Mouse monoclonal antibody against FLAG-tag (MBL, Nagoya, Japan), HRP-conjugated goat anti-mouse IgG (Sangon Biotech, Shanghai, China), and GAPDH monoclonal antibody (Bioworld Technology, Nanjing, China) were used in Western blot analysis. Alexa Fluor^® 488-conjugated goat anti-mouse IgG antibody (Jackson ImmunoResearch, West Grove, PA, USA) was used as the secondary antibody in the immunofluorescence assay.