Tween 20

Proteins were separated by 10% SDS-PAGE and then transferred to a polyvinylidene difluoride (PVDF) membrane. After blocking with 5% skim milk in Tris-buffered saline (137 mM NaCl, 20 mM Tris–HCl, pH8.0) containing 0.05% Tween-20 (Sangon Biotech, Shanghai, China), the membrane was incubated with a primary antibody (Abcam, Cambridge, UK) overnight at 4 °C. Subsequently, the membrane was incubated with HRP (horseradish peroxidase)-conjugated secondary antibody (Abcam) at 37 °C for 2 h, followed by examination with enhanced chemiluminescence system (ChampGel, Beijing, China).

All oligonucleotides (HPLC purified) were synthesized and modified by Sangon Biotechnology Co., Ltd. (Shanghai, China). D-PBS buffer, 20× PBS buffer, 5× TBE buffer, RNase-free ddH₂O, Tween-20, Bovine Serum Albumin (BSA), HRP-labeled Streptavidin (SA-HRP), and EL-TMB Chromogenic Reagent kit were purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). ASFV p30 recombinant protein, p30 monoclonal antibody, porcine reproductive and respiratory syndrome virus (PRRSV) recombinant N protein, staphylococcus aureus enterotoxin type A recombinant protein (SEA), and pig immunoglobulin G (IgG) were purchased from Aiyi Biotechnology. (Nanjing, China). 6× His tag peptides were bought from Synpeptide Biotechnology Co., Ltd. (Nanjing, China). Swine serum was sourced from Solarbio Science & Technology Co., Ltd. (Beijing, China). NHS (N-hydroxysuccinimide)-Activated Magnetic Beads (NHS-MB) were purchased from Beaver Biosciences Inc. (Suzhou, China). Taq PCR Master Mix and ChamQ Universal SYBR qPCR Master Mix were purchased from Vazyme Biotechnology Co., Ltd. (Nanjing, China). The 96-well polystyrene microtiter plate (MaxiSorp) was bought from Thermo Fisher Scientific (Shanghai, China). ASFV-positive freeze-dried serum, porcine negative serum national reference was purchased from the National Center of Veterinary Culture collection (CVCC) (Beijing, China).

The PCR Master Mix Assay kit and the DL500 DNA marker were purchased from Takara Bio Inc. (Dalian, China). GoldView-I nucleic acid stain, streptavidin, PEG-20000, and sodium chloride were obtained from Solarbio Science & Technology Co. Ltd. (Beijing, China). Tris-(hydroxymethyl) aminomethane (Tris), HAuCl₄·3H₂O, and Tween-20 were bought from Sangon Biotechnology Inc. (Shanghai, China). A MagIso DNA/RNA isolation kit was obtained from Xi'an Tianlong Science and Technology Co. Ltd. (Xian, China). A sample pad, a conjugate pad, absorbent paper, and a PVC baseboard were obtained from Jie Yi Biotechnology Co. Ltd. (Shanghai, China). UniSart CN 140 nitrocellulose membranes were obtained from Millipore (Billerica, MA, USA). Synthetic oligonucleotides were obtained from Tsingke Biotechnology Co. Ltd. (Beijing, China).