Thymidine → Aflatoxin M1

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Reaktionsschritte

Materials: All inorganic chemicals and organic solvents were of reagent grade or better. Bovine serum albumin (BSA) (fatty acid free and fraction V) ovalbumin (OA) (crude and fraction VII), polyethylene sorbitan monolaurate (Tween 20), 2,2'-azino-di-(3-ethylbentzthiazoline)sulfonic acid (ABTS), hydrogen peroxide dicyclohexylcarbodiimide, N-hydroxysuccidimide, dimethylformamide (DMF), polyethylene glycol (PEG) (MW 1450), insulin, oxaloacetate, hypoxanthine (H), amino-pterin (A), thymidine (T) and 2,6,10,14 tetramethylpentadecane (pristane), horseradish peroxidase (HRP) Type VI were purchased from Sigma Chemical Company (St. Louis, Mo.); tetrahydrofuran from Aldrich Chemical Company (Milwaukee, Wis.); Freund's complete and incomplete adjuvants from Difco (Detroit, Mich.); goat anti-mouse IgG conjugated to horseradish peroxidase from Cooper Biomedical (Malvern, Pa.); Dulbecco's Modified Eagle's Medium (DMEM), Penicillin/Streptomycin Solution (Pen/Strep) (100,000 U/ml), NCTC Medium 135, fetal bovine serum (FBS), and sodium pyruvate from Gibco Laboratories (Grand Island, N.Y.); microculture plates (96- and 24-well plates) from Costar (Cambridge, Mass.); microtiter plates from NUNC, Vangard International (Neptune, N.J.). Aflatoxins B1, B2, G1, G2 were purchased from Sigma Chemical Company (St. Louis, Mo.); aflatoxin M1 was generously provided by the Department of Botany and Plant Pathology, Michigan State University, East Lansing, Mich. Subclass identification kit was purchased from Boehringer Mannheim Biochemicals (Indianapolis, Ind.). The myeloma cell line P3/NS 1/1-Ag-4 (NS-1) (ATCC TIB 18) was purchased from The American Type Culture Collection (Rockville, Md.). Mice were purchased from Charles River Laboratories (Wilmington, Mass.).

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