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2 protocols using polyethylene glycol peg 1450

1

Monoclonal Antibody Generation and Characterization

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DA, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), methanol, polyethylene glycol 1450 (PEG-1450), phosphate-buffered saline (PBS, pH 7.4), borate buffer (0.085 M), ovalbumin (OVA), bovine serum albumin (BSA), Freund’s complete and incomplete adjuvant, pristane (2,6,10,14-Tetramethylpentadecane), HAT medium, horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody, polyethylene glycol (PEG-1450), IgG1, IgG2a, IgG3, IgM, IgA mouse mAb isotyping kit, hypoxanthine-thymidine (HT) medium, and RPMI-1640 were acquired from Sigma-Aldrich Chemical (St. Louis, MO, USA). The murine myeloma cell line Sp2/0 was stocked in liquid nitrogen at our laboratory. All Balb/c mice (female, six to eight-weeks old) used in the present study were obtained from the Wushi animal laboratory (Shanghai, China). All other chemical reagents used in the current study were acquired commercially in China and were of chemical grade.
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2

Mycotoxin Detection Assay Development

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Citrinin (CIT), Aflatoxin B1 (AFB1), Fumonisin B1 (FB1), Patulin (PAT), Ochratoxin A (OTA), polyethylene glycol (PEG) 1450, hypoxanthine aminopterin and thymidine (HAT) medium, hypoxanthine-thymidine (HT) medium, complete and incomplete Freund adjuvant were purchased from Sigma (USA). The mouse McAb isotyping kit was purchased from Hycult Biotechnology (Netherlands). The SP2/0 myeloma cell line was stored in our lab. The Ridascreen Fast CIT ELISA kit was purchased from the R-Biopharm group (Germany). Goat anti-mouse IgG (H+L) was purchased from BioWorld (USA). Protein-G Sepharose Fast Flow Columns were purchased from GE Healthcare (USA). Chloroauricacid (HAuCl4) and trisodium citrate were obtained from Shanghai Chemical Reagents (Shanghai, China). The nitrocellulose membranes (Millipore 135, Millipore 180) were purchased from Millipore (USA). Othertypes of nitrocellulose membranes (Sartorius CN 140, PALL Vivid 170), conjugation pads (GL0194, Ahlstrom 8964, Ahlstrom 6613), sample pads (GL-b01, GL-b02) and absorbent pads were purchased from Jieyi Bio. Co. (China).
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