Mouse anti his monoclonal antibody

Different positive sera were used to detect the specificity of ELISA. Classical swine fever virus (CSFV)-, porcine circovirus type 2 (PCV2)-, porcine reproductive and respiratory syndrome virus (PRRSV)-positive serum samples, and PEDV reference positive and negative sera were prepared and kept by our lab. Transmissible gastroenteritis virus (TGEV)-positive serum was kindly provided by Hunan University.
A total of 213 pig blood samples were collected from Aug 2013 to Apr 2019 from local farms in Zhejiang Province. Serum was separated from blood samples by centrifugation at 1500 × g for 10 min and kept at -20 °C until use.
Previously produced mouse anti-S1 monoclonal antibody 6B4/6G1/1C11 could recognize S1T1, S1T2, S1T3, respectively. PEDV positive serum and mouse anti-His monoclonal antibody (Beyotime, China) were used as primary antibody. HRP-labeled goat anti-mouse IgG antibody (Beyotime, China). HRP-labeled goat anti-pig IgG antibody (Lifetechnology, USA), Alexa Fluor 488-conjugated goat anti-pig IgA antibody (Lifetechnology, USA) and Alexa Fluor 568-conjugated goat anti-mouse IgG antibody (Lifetechnology, USA) were used as secondary antibody.

The rabbit anti-NP polyclonal antibody and mouse anti-M1 monoclonal antibody were generated as previously described [71 (link)]. The rabbit anti-NS1 polyclonal antibody was generated as previously described [72 (link)]. The mouse anti-FLAG M2 monoclonal antibody (Sigma-Aldrich, USA), mouse anti-Myc monoclonal antibody (Santa Cruz Biotechnology, USA), mouse anti-His monoclonal antibody (Beyotime, China), rabbit anti-IAV PB1 polyclonal antibody (GeneTex, USA), rabbit anti-IAV PB2 polyclonal antibody (GeneTex, USA), mouse anti-Lamin B1 monoclonal antibody (Santa Cruz Biotechnology, USA), mouse anti-β-actin monoclonal antibody (TransGen Biotech, China), mouse anti-GAPDH monoclonal antibody (Abcam, UK), HRP-conjugated goat anti-mouse IgG (Beyotime, China), HRP-conjugated goat anti-rabbit IgG (Beyotime, China), HRP-conjugated rabbit anti-mouse IgG (Cell Signaling Technology, USA), and HRP-conjugated mouse anti-rabbit IgG (Cell Signaling Technology, USA) were purchased from the indicated companies. Poly(I:C) sodium salt (Sigma, USA), human IFN-β (PeproTech, USA), RNase R (Lucigen, USA), RiboLock RNase Inhibitor (Thermo Fisher Scientific, USA), protease inhibitor cocktail (Roche, Switzerland), isopropyl-β-D-thiogalactopyranoside (IPTG; Calbiochem, USA), and tolylsulfonyl phenylalanyl chloromethyl ketone (TPCK)-trypsin (Sigma-Aldrich, USA) were purchased from the indicated companies.