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3 protocols using carbaryl

1

Cholinesterase Inhibitor Assay Protocol

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S-butyrylthiocholione iodide (BTC), 5-5´-dithiobis-(2-nitrobenzoic acid) (DTNB), propionylthiocholine iodide (PTC), aldicarb (98% purity), carbofuran (99.5% purity), carbaryl (99.5% purity), and propoxure (99.6%) were purchased from Wako Pure Chemical Industries (
www.wako-chem.co.jp). Phosphamidon and monocrotophos with 99.9% purity were purchased from Accustandard (
www.accustandard.com). Acetylcholine iodide (ATC), eserine, tris (hydroxymethyl) aminomethane, Triton X-100, and bovine serum albumin were obtained from Sigma-Aldrich (
www.sigmaaldrich.com).
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2

Apoptosis Assay of Pesticide-Treated Cells

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RPMI 1640 medium was purchased from GIBCO (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from JRH Biosciences (Lenexa, KS, USA), and heat-inactivated at 56 °C for 30 min prior to use. Glutamine, 2-mercaptoethanol (2-ME) and propidium iodide (PI) were obtained from Sigma (St. Louis, MO, USA). Fluorescein isothiocynate (FITC)-anti human annexin V and FITC-anti human active caspase-3, Z-DEVD-FMK (a caspase-3 inhibitor), Z-VAD-FMK (a general caspase inhibitor), Z-FA-FMK (a negative control for Z-DEVD-FMK and Z-VAD-FMK), and Cytofix/cytoperm solution were purchased from BD Pharmingen (San Diego, CA, USA). Carbaryl, maneb, thiram and ziram were obtained from Wako Pure Chemical Industries (Osaka, Japan) and prepared as stock solutions in DMSO.
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3

Analytical Workflow for Xenobiotic Metabolites

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Methiocarb, carbaryl, 1-naphthol, methiocarb sulfoxide, methiocarb sulfone, 4-methylthio-3,5-xylenol (MX), 3,5-dimethyl-4-(methylsulfinyl)phenol (SP), 3,5-dimethyl-4-(methylsulfonyl)phenol (SOP), menadione, 2-hydroxypyrimidine, rat albumin and human albumin were purchased from Wako Pure Chemical Industries (Osaka, Japan). Eserine, 2-chloro-3,4-dimethoxybenzyl (CDMB), bis(4-nitrophenyl)phosphate (BNPP), 5-pregnen-3β-ol-20-one-16α-carbonitrile (PCN), artemisinin, and bezafibrate (BZF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Infinity pure dimethyl sulfoxide (DMSO; > 99.5% pure) was purchased from Wako Pure Chemical Industries. Human recombinant CYP and FMO isoforms expressed in a baculovirus system and pooled human plasma were purchased from BD Gentest (Woburn, MA, USA).
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