Afb1 bsa

AFB₁–BSA, rabbit anti-AFB₁, goat anti-rabbit IgG horseradish peroxidase (HRP) conjugate, 3,3′,5,5′-tetramethylbenzidine (TMB) substrate, carbonate–bicarbonate buffer (capsule), multi-walled carbon nanotubes (MWCNTs), medium molecular weight chitosan (CS) and potassium ferricyanide, K₃[Fe(CN)₆], were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Aflatoxin B₁ (AFB₁) standard solution was purchased from Supelco Analytical (Bellefonte, PA, USA). Skimmed milk powder was obtained from Nacalai Tesque (Kyoto, Japan). Other reagents were of analytical grade, and all aqueous solutions were prepared using deionized water. Phosphate buffer saline (PBS) was prepared at 10× stock concentration and diluted to 1×. Tween 20 (0.05%) was added to 1 L of 1× PBS to make PBST as a washing buffer in ELISA. For supporting electrolyte, 0.17 g of potassium ferricyanide, K₃[Fe(CN)₆], was added into 100 mL of 1× PBS to make a 5 mM solution, to study the reversible electrochemical behavior in cyclic voltammetry analysis.

AFB₁-BSA, rabbit anti-AFB₁, goat anti-rabbit IgG horseradish peroxidase (HRP) conjugate, 3,3′,5,5′-tetramethylbenzidine (TMB) substrate, carbonate-bicarbonate buffer (capsule), multi-walled carbon nanotubes (MWCNTs), medium molecular weight chitosan (CS) and potassium ferricyanide, K₃[Fe(CN)₆], were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aflatoxin B₁ (AFB₁) standard solution (20 µg/mL in methanol) was purchased from Supelco Analytical (Bellefonte, PA, USA). Skimmed-milk powder was obtained from Nacalai Tesque (Kyoto, Japan). Other reagents were of analytical grade, and all aqueous solutions were prepared using deionized water. Phosphate buffer saline (PBS) was prepared at 10× stock concentration and diluted to 1×. Tween 20 (0.05%) was added to 1 L of 1× PBS to make PBST as a washing buffer in ELISA. For supporting electrolyte, 0.17 g K₃[Fe(CN)₆] was added into 100 mL of 1× PBS to make a 5 mM solution.

BSA, OVA, Freund’s complete/incomplete adjuvant, HEPES, L-Glu, HT/HAT (50×), PEG1450, and DMSO (bio-sterile grade) were purchased from Sigma, (St. Louis, MO, USA). AFB1, AFB2, AFM1, AFM2, AFG1, and AFG2 were purchased from Tianjin Alta. Double antibodies (penicillin) (100×), non-essential (100×), were purchased from Thermofisher, (Waltham, MA, USA). RPMI-1640 basal medium was purchased from Cytiva, (Beijing, China). Growth factors were purchased from Beijing Biodragon Immunotechnologies Co., Ltd, (Beijing, China). FBS was purchased from Zhejiang Tianhang Biotechnology, (Zhejiang Province, China). Goat anti-mouse IgG-HRP (2 mg/mL) was purchased from Shandong Lvdu Bio-science & technology CO., Ltd (Binzhou, China). AFB1-BSA was purchased from sigma, BeijingBoaolong Immunotech, USA, and provided by the Research Institute of China Agricultural University, respectively. The anti-AFB1 monoclonal antibody 1c11 was provided by the Research Institute of China Agricultural University and purchased from Shandong Lvdu Bio-science & technology Co., Ltd. The mouse myeloma cell line is SP2/0 provided by Shandong Fenghua Biological Co., Ltd, (Zibo, China). All other reagents were analytically pure. The 96-well polystyrene enzyme plates (3590) and cell culture plates (3599) were purchased from Costar, (New York, NY, USA).