A transfected Chinese hamster ovary (CHO), Hela, a murine interleukin-3 dependent pro-B (Ba/F3), PC12, and rat basophil leukemia cell lines were obtained from Eurofins Scientific (Eurofins-Cerep, Le Bois I’Eveque, France). Buffers—Dulbecco’s modified Eagle medium (DMEM) buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, and Hank’s balanced salt solution (HBSS) buffer—were purchased from Invitrogen (Carlsbad, CA, USA). The reference agonists: 5′-N-ethylcarboxamidoadenosine (NECA), epinephrine bitartrate, epinephrine, DPDPE, CP 55940, linoleic acid, GLP-1(7–37, arginine vasopressin (AVP), and serotonin, and antagonists: ZM 241385, RX-821002, rauwolscine, naltriben mesylate, AM 281, exendin-3(9–39), [d(CH2)51, Tyr (Me)2]-AVP, (S)-WAY-100635, and GR55562) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals and reagents purchased from Merck and Fluka were of the highest available grade unless otherwise stated.
Am281
Manufactured by Merck Group
Sourced in United States, Italy
AM281 is a synthetic cannabinoid receptor antagonist. It is commonly used in research applications involving the endocannabinoid system.
Automatically generated - may contain errors
Lab products found in correlation
Win 55 212 2, by Merck Group (3 mentions)
Epinephrine, by Merck Group (1 mentions)
Dulbecco s modified eagle medium dmem buffer, by Thermo Fisher Scientific (1 mentions)
Glp 1 7 37, by Merck Group (1 mentions)
Rx 821002, by Merck Group (1 mentions)
S way 100635, by Merck Group (1 mentions)
Cp55940, by Merck Group (1 mentions)
Linoleic acid, by Merck Group (1 mentions)
4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes buffer, by Thermo Fisher Scientific (1 mentions)
5 n ethylcarboxamidoadenosine neca, by Merck Group (1 mentions)
Hank s balanced salt solution hbss buffer, by Thermo Fisher Scientific (1 mentions)
Epinephrine bitartrate, by Merck Group (1 mentions)
Rauwolscine, by Merck Group (1 mentions)
Zm241385, by Merck Group (1 mentions)
Arginine vasopressin avp, by Merck Group (1 mentions)
Dpdpe, by Merck Group (1 mentions)
Dmem f12 medium, by Merck Group (1 mentions)
Methyl viologen dichloride hydrate, by Merck Group (1 mentions)
Penicillin streptomycin solution hybri max p s, by Merck Group (1 mentions)
Fetal bovine serum l glutamine l glut, by Merck Group (1 mentions)
7 protocols using am281
1
Cell Lines and Reagents for Bioassays
2
Oxidative Stress and Cannabinoid Signaling
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(±)-α-tocopherol; methyl viologen dichloride hydrate; DMEM:F12 medium; penicillin–streptomycin solution hybri-max (P/S); fetal bovine serum (FBS); L-glutamine (L-Glut); non-essential amino acid solution (NEAA); trypan blue; AM281; WIN55,212-2; mesylate salt; 2-mercaptobenzothiazole; and 1,5-diaminophtalene were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Methanol and acetonitrile were purchased from Panreac (Barcelona, Spain) and cesium chloride was purchased from HoneyWell (Charlotte, NC, USA).
3
Acetaldehyde-Induced Liver Injury Protocol
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Acetaldehyde 99.98% (Sigma-Aldrich, Milan, Italy) was daily diluted with tap water to a final concentration of 8% v/v; each intragastric ACD administration provided 450 mg/kg. The selective CB1 receptor antagonist AM281 (Sigma-Aldrich, Milan, Italy) was suspended in saline solution containing 3% Tween 80 and administered i.p. at 2.5 mg/kg, in ACD and CTR animals at day 5, 3 and 12 h after the last intragastric administration [modified from Ref. (59 (link))]. The dose of AM281 used in this study was chosen to avoid any aspecific effect (60 (link), 61 (link)).
4
Cannabinoid Modulation of Contextual Fear
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7-NI (15, 30, and 60mg/kg, Sigma-Aldrich), a preferential nNOS inhibitor, was dissolved in 5% Tween 80 in NaCl 0.9%; WIN55,212-2 (Win; 0.1, 0.3, and 1.0mg/kg, Sigma-Aldrich), a nonselective cannabinoid agonist, and AM281 (1, 2, and 4mg/kg), a potent and selective CB1 antagonist, were dissolved in 10% dimethylsulfoxide (DMSO) in NaCl 0.9% and administered i.p. to WT mice 30 minutes before the first reexposure to the context chamber (Maren, 1998; Rutkowska et al., 2006; Gilhotra and Dhingra, 2009 ; Gomes et al., 2011 ). URB597 (URB; 0.3, 1, and 3mg/kg), an inhibitor of the FAAH enzyme that metabolizes the ECB anandamide, was dissolved in 10% DMSO in NaCl 0.9% and administered i.p. to WT mice 1 hour before the first reexposure to the context chamber (Gomes et al., 2011 ). All drugs were administered in a fixed volume of 10mL/kg of body weight. The animals were also reexposed to the same context 48, 72, and 96 hours after the first chamber exposure.
For evaluation of freezing behavior, independent groups of WT and iNOS KO mice received i.p. injections of 7-NI (effective dose, 30mg/kg) or URB (effective dose, 3.0mg/kg) 30 minutes or 1 hour, respectively, before the first reexposure to the context chamber previously paired with electric footshocks.
For evaluation of freezing behavior, independent groups of WT and iNOS KO mice received i.p. injections of 7-NI (effective dose, 30mg/kg) or URB (effective dose, 3.0mg/kg) 30 minutes or 1 hour, respectively, before the first reexposure to the context chamber previously paired with electric footshocks.
5
Pharmacological Modulation of Synaptic Transmission
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All drugs were diluted in ACSF to the appropriate concentration. In the experiments presented here the drug used were: the sodium channel blocker tetrodotoxin (TTX; 0.1 μM); the NMDA receptor antagonist D -(-)-2-amino-5-phosphonopentanoic acid (APV, 50 μM; Tocris); the AMPA/kainate receptor antagonist (DNQX, 20 μM; Sigma); the synthetic CB receptor agonist (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone mesylate (WIN 55,212-2; 1 μM); the endogenous CB agonist N-(2-hydroxyethyl)-5Z,8Z,11Z,14Z-eicosatetraenamide (Anandamide, 1 μM); the CB1 receptor antagonist 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4-morpholinyl-1H-pyrazole-3-carbox-amide (AM281, 1 μM); the selective vanilloid receptor antagonist N-[2-(4-chlorophenyl)ethyl]-1,3,4,5-tetrahydro-7,8-dihydroxy-2H-2-benzazepine-2-carbothioamide (Capsazepine, 5 μM); the opioid receptor agonist DAMGO (0.1 μM); the neurotrophin brain-derived neurotrophic factor (BDNF; 20 ng/ml); the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid (BAPTA). All chemicals were obtained from Tocris.
6
Cannabinoid Receptor Modulation in Neuroscience
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HU210 and AM281 were purchased from Sigma (USA) and dissolved in vehicle of 2:1:37 of dimethyl sulfoxide (DMSO): Tween-80: 0.9% saline. All drugs and vehicle were dispensed on the day of experiments. HU210 was acutely administrated an hour before the behavioral or electrophysiological experiments by intraperitoneal injection.
Tetrodotoxin (TTX) and picrotoxin (PTX) were purchased from Sigma (USA), which were made into concentrated stock solutions and diluted in artificial cerebrospinal fluid (ACSF) to the final concentration on the day of testing, and continuously poured into the recording chamber. CNO (purchased from BrainVTA, China) was pre-dissolved in DMSO and then diluted with 0.9% saline to a final concentration of 0.5% just before the experiments. For drug stocks prepared with DMSO, the final DMSO concentration was less than 0.1%.
Tetrodotoxin (TTX) and picrotoxin (PTX) were purchased from Sigma (USA), which were made into concentrated stock solutions and diluted in artificial cerebrospinal fluid (ACSF) to the final concentration on the day of testing, and continuously poured into the recording chamber. CNO (purchased from BrainVTA, China) was pre-dissolved in DMSO and then diluted with 0.9% saline to a final concentration of 0.5% just before the experiments. For drug stocks prepared with DMSO, the final DMSO concentration was less than 0.1%.
7
Cannabinoid Effects on Reward Learning
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The cannabinoid agonist WIN 55, 212-2 mesylate (WIN; Tocris Bioscience, UK) and the CB1 receptor antagonist AM281 (Sigma, Italy) were initially dissolved in 3% ethanol; followed by 3% Tween 20 and 94% saline (Cannizzaro et al., 2016) . WIN was administered intraperitoneally (i.p.) at a dose of 1 mg/kg (volume of 1 mL/kg), 30 min before the cued reward-conditioned learning session (pre-conditioning WIN) or 30 min before the EOR experiment (post-conditioning WIN). WIN dosage was chosen on the basis of previous studies (Brancato et al., 2016; Darmani, 2001; Järbe, 2006) , in order to avoid unspecific effects due to neural and locomotor impairment (Rizzo et al., 2014) . AM281 was administered i.p. at a dose of 2 mg/kg, 30 min before WIN (Suenaga and Ichitani, 2008) . Control rats received the same volume of vehicle at the same time points (1 mL/kg).
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