Goat anti rabbit ig hrp conjugate secondary antibody

The anti-FLAG mouse monoclonal antibody (#F3165), anti-pTurboGFP rabbit polyclonal antibody (#AB513) and anti-GAPDH mouse monoclonal antibody (#60004-1-Ig) were purchased from Sigma-Aldrich, Everogen and Proteintech, respectively. To obtain these antibodies against NS80, NS38, VP3 and VP5 proteins of GCRV, the 2~160 amino acids (aa) of NS38, 500~692 aa of NS80, 2~200 aa of VP3 or 451~ 648 aa of VP5 was inserted into the pET-32a (+) vector (EMD Millipore) for prokaryotic expression. Purified recombinant proteins were used to immunize Japanese White rabbits to acquire the rabbit polyclonal antibodies and mice to acquire the mouse polyclonal antibodies. The antiserum from the rabbit was affinity-purified on antigen-coupled CNBr-activated agarose (GE Healthcare). Goat-anti-mouse Ig-HRP conjugate secondary antibody, Goat-anti-rabbit Ig-HRP conjugate secondary antibody, Alexa Fluor 488-conjugated secondary Ab against mouse IgG, Alexa Fluor 594-conjugated secondary Ab against rabbit IgG, 6-diamidino-2-phenylindole (DAPI), Lipofectamine 2000 and Protease inhibitor cocktail were purchased from Thermo Fisher Scientific. Dimethylsulfoxide (DMSO) and the FLAG^® Immunoprecipitation Kit was purchased from Sigma-Aldrich. Brefeldin A (BFA, S7046) was purchased from Selleck. Golgi-tracker red (C1043) was purchased from Beyotime.

The anti-FLAG mouse monoclonal antibody (number F3165), anti-pTurboGFP rabbit polyclonal antibody (number AB513), and anti-GAPDH mouse monoclonal antibody (number 60004-1-Ig) were purchased from Sigma-Aldrich, Everogen, and Proteintech, respectively. The anti-NS38 polyclonal mouse antibody and anti-NS80 polyclonal rabbit antibody against the GCRV-873 strain were prepared previously (7 (link)– (link)10 (link)). Lovastatin (S2061), VitD (S1466), MG132 (S2619), and 3-MA (S2767) were purchased from Selleck (Shanghai, China). Goat-anti-mouse immunoglobulin-horseradish peroxidase (Ig-HRP) conjugate secondary antibody, goat-anti-rabbit Ig-HRP conjugate secondary antibody, Alexa Fluor 488-conjugated secondary Ab against mouse IgG, Alexa Fluor 594-conjugated secondary Ab against rabbit IgG, 4′,6-diamidino-2-phenylindole (DAPI), Subcellular Protein Fractionation kit, Lipofectamine 2000, and protease inhibitor cocktail were purchased from Thermo Fisher Scientific. The FLAG immunoprecipitation kit and chloroquine (C6628) were purchased from Sigma-Aldrich.