Rabbit monoclonal gfp antibody

Cells were lysed in RIPA lysis buffer [150 mM NaCl, 1% Non-ionic detergent, 1% Sodium deoxycholate, 0.1%SDS, (Biomax, Republic of Korea)]. About 20 μg of proteins per sample were separated by SDS-PAGE using a 10%polyacrylamide gels and transferred to nitrocellulose membranes (Cytiva, USA). Membranes were blocked with 10% (w/v) non-fat dry milk in TBST buffer [100 mM Tris (pH 7.6), 150 mM NaCl, 0.1% (w/v) Tween-20] for 1 h at room temperature. Membranes were incubated with a 1:1,000 dilution of rabbit polyclonal anti-Gag antibody, 1:500 dilution of rabbit polyclonal anti-Env antibody, 1:5,000 dilution of rabbit monoclonal GFP antibody (Abcam, UK), and 1:5,000 dilution of rabbit monoclonal β-actin antibody (Cell Signaling Technology, USA) at 4°C overnight. The membranes were then incubated with a 1:10,000 dilution of secondary antibody, goat anti-rabbit IgG conjugated to HRP (BioRad, USA), for 1 h at room temperature. The chemiluminescent signal was visualized by exposing the blots to a chemiluminescence imaging system, E-blot (e-BLOT Life Science, China). Rabbit polyclonal anti-Env antibody was designed to be specific for PFV Env-SU domain and was custom-produced by AbClon (Korea).