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2 protocols using methyllycaconitine mla citrate salt hydrate

1

Neurotransmitter Receptor Binding Assays

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Methanol, hexane, chloroform, ethyl acetate, butanol, acetonitrile, (−)-nicotine (NIC), methyllycaconitine (MLA) citrate salt hydrate, mecamylamine (MEC) hydrochloride, nomifensine, (−)-lobeline hydrochloride, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma Aldrich (St. Louis, MO, USA). Streptomycin (10,000 μg/ml), penicillin (10,000 units/ml), fetal bovine serum (FBS), and Dulbecco’s Modified Eagle Medium (DMEM) were purchased from Life Technologies Corporation (Grand Island, NY, USA). Quercetin was purchased from Chromadex (Irvine, CA, USA). [3H]-epibatidine (S.A. = 30 Ci/mmol), [3H]-cytisine (S.A. = 16 Ci/mmol), [3H]-MLA (S.A. = 60 Ci/mmol), [3H]-GBR12935 (S.A. = 40 Ci/mmol), 45CaCl2 (S.A. = 12.05 mCi/mg), and [3H]-DA (S.A. = 60 Ci/mmol) were purchased from American Radiolabeled Chemicals, Inc. (St. Louis, MO, USA). All other chemicals and materials were purchased from Fisher Scientific (Pittsburgh, PA, USA), unless otherwise stated.
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2

NADPH Oxidase Inhibition Protocol

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The NADPH oxidase inhibitor diphenyleneiodonium (DPI; SIGMA) was dissolved in DMSO at 10 mM, and the solutions were diluted into the control medium to prepare working solution (1 μM). The concentration of DMSO in the medium was 0.01 %. As control for DPI application, the same amount of DMSO (0.01 %) was added as vehicle. (−)-Nicotine hydrogen tartrate salt, α-bungarotoxin-tetramethylrhodamine (α-Bgt) and methyllycaconitine (MLA) citrate salt hydrate were purchased from SIGMA.
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