Nicotine [(−)-nicotine hydrogen tartrate] was purchased from Sigma Chemical Company (St. Louis, MO, USA). Nicotine was diluted in saline, and the pH of the solution was adjusted to 7.0 with dilute NaOH. Cocaine [(−)-cocaine hydrochloride] was obtained from NIDA, NIH (Rockville, MD) and diluted in saline. The dose of nicotine and cocaine (both 30 μg/kg/injection) for this study was selected based on our previous studies in which this dose maintained maximal rates of responding. The mGluR2/3 agonist LY379268[(1R,4R,5S, 6R)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate] was provided by Dr. Athina Markou (University of California San Diego; custom synthesized by ANAWA) and dissolved in sterile water. LY379268 or the vehicle was always injected i.m. 30 min before the session.
Nicotine nicotine hydrogen tartrate
Manufactured by Merck Group
Sourced in United States
Nicotine [(−)-nicotine hydrogen tartrate] is a laboratory reagent used in chemical and biological research. It is the hydrogen tartrate salt of the naturally occurring alkaloid nicotine. This product is intended for use in research and development applications, and its core function is to serve as a chemical standard or reference material for analysis and experimentation purposes.
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Lab products found in correlation
Baclofen, by Merck Group (1 mentions)
Elisa assay kits, by Enzo Life Sciences (1 mentions)
Elisa assay kits for, by Thermo Fisher Scientific (1 mentions)
Hemicholinium 3 hc 3, by Merck Group (1 mentions)
Wortmannin, by Bio-Techne (1 mentions)
Pik 75, by Cayman Chemical (1 mentions)
Pi 828, by Bio-Techne (1 mentions)
Gsk 690693, by Bio-Techne (1 mentions)
Lestaurtinib, by Bio-Techne (1 mentions)
As 605240, by Cayman Chemical (1 mentions)
Human tnf α, by Thermo Fisher Scientific (1 mentions)
Cucurbitacin 1, by Bio-Techne (1 mentions)
Ly294002, by Bio-Techne (1 mentions)
Ouabain, by Merck Group (1 mentions)
Bodipy conjugated ouabain, by Thermo Fisher Scientific (1 mentions)
5 protocols using nicotine nicotine hydrogen tartrate
1
Nicotine and Cocaine Addiction Modulation
2
Nicotine and AZD8529 Solution Preparation
3
Gemcitabine Signaling Pathway Exploration
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Gemcitabine (Gemzar hydrochloride) was purchased from Eli Lilly (Indianapolis, IN, USA). Baclofen, GABA and nicotine ((-)-Nicotine hydrogen tartrate) were purchased from Sigma Aldrich (St. Louis, MO, USA). Antibodies against the following proteins were purchased from Abcam (Cambridge, MA, USA): GABA-B receptor 1, GABA-B-receptor 2, beta-actin, metalloproteinase-2 (MMP-2), metalloproteinase-9 (MMP-9) and early growth response gene 1 (EGR-1). ELISA assay kits for the quantitative determination of intracellular cyclic adenosine monophosphate (cAMP) were purchased at Enzo Life Sciences (Farmingdale, NY, USA). ELISA assay kits for the quantitative determination of the phosphorylated signaling proteins extra cellular signal regulated kinase (ERK), cAMP response element binding protein (CREB), v-akt murine thymoma viral oncogene homolog (AKT), for Caspase-3 and for v-src avian sarcoma viral oncogene homolog (Src) were purchased at Invitrogen (Carlsbad, CA, USA) and MBL International Corporation (Woburn, MA, USA), respectively.
4
Characterization of HEK293 Cells Expressing Nicotinic Receptors
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The HEK293 (293) cell line stably co-transfected with rat nAChR subunits α4 and β2 are well characterized [6 (link),11 (link),12 (link)], and they were maintained as described previously [9 (link),10 (link),14 (link)]. This cell line also expresses TNFR1 (only weak expression of TNFR2 is detected) and no additional human nAChR subunits or acetylcholine synthetic enzymes are detected [9 (link),10 (link)]. As before treatments were conducted 48 hours after cell plating [9 (link),10 (link),14 (link)]. The source and optimal concentrations of the inhibitors used are as follows: From Sigma; 1 μM nicotine (nicotine hydrogen tartrate); 250 μM choline (choline chloride); hemicholinium-3 (HC3; 50 μM). From Biosource; 25 ng/ml human TNFα. From Tocris Bioscience: 1,2,3,4,5,6-Hexabromocyclohexane (1-6Hex; 50 μM), tyrphostin 2-cyano-3-(3,4-dihydrophenyl)-N-(phenylmethyl)-2-propenamide (AG-490; dose as in text), Cucurbitacin I (100 nM); GSK690693 (5 μM), Lestaurtinib (300 nM), Ly294002 (10 μM), PI 828 (20 nM), SB2020190 (10 μM), Wortmannin (3 μM), ZM 39923 (50 μM), ZM449829 (50 μM). From Cayman Chemical: AS-605240 (20–100 nM); PIK-75 (50 μM). Most inhibitors were dissolved in either DMSO or ethanol as the carrier and in all cases the carrier was added to cultures at the same concentration as before [9 (link),10 (link),14 (link)].
5
Fluorescent Labeling of Ouabain
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Ouabain was obtained from Sigma-Aldrich. Rhodamine-conjugated α-bungarotoxin was from Biotium, Inc., and BODIPY-conjugated Ouabain was from Invitrogen. Nicotine ((−)nicotine hydrogen tartrate) and other chemicals were from Sigma-Aldrich.
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