Aripiprazole, quinpirole, and haloperidol were purchased from Tocris (Ellisville, MO, USA). A769662 was obtained from LC Laboratories (Woburn, MA, USA). Thioridazine was obtained from Cayman Chemical (Ann Arbor, MI, USA). Irradiation was performed at room temperature by using a 137Cs gamma‐ray source Gammacell 3000 manufactured by Nordion (Ottawa, ON, Canada).
Haloperidol
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
Haloperidol is a laboratory reagent manufactured by Bio-Techne. It is a neuroleptic medication commonly used in research applications. The core function of Haloperidol is to act as a potent dopamine D2 receptor antagonist, which can be used to study dopaminergic signaling pathways and related biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Clozapine, by Bio-Techne (6 mentions)
Olanzapine, by Bio-Techne (3 mentions)
Aripiprazole, by Bio-Techne (2 mentions)
Pentazocine, by Merck Group (2 mentions)
Pd 144418, by Bio-Techne (2 mentions)
Sch23390, by Bio-Techne (2 mentions)
3h pentazocine, by PerkinElmer (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Haloperidol, by Merck Group (2 mentions)
Quinpirole, by Bio-Techne (1 mentions)
Thioridazine, by Cayman Chemical (1 mentions)
A 769662, by LC Laboratories (1 mentions)
Gammacell 3000, by Nordion (1 mentions)
Bkm120, by Merck Group (1 mentions)
L 741 626, by Bio-Techne (1 mentions)
Trichostatin a, by Cayman Chemical (1 mentions)
Gallein, by Santa Cruz Biotechnology (1 mentions)
Human brain cerebral cortex protein medley, by Takara Bio (1 mentions)
L 745870, by Bio-Techne (1 mentions)
Clozapine n oxide, by Bio-Techne (1 mentions)
20 protocols using haloperidol
1
Pharmacological Treatments and Irradiation
2
BRET Quantification of Ligand Binding
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Drug-induced BRET is conducted as optimized previously (i.e., transfection and experimental conditions) (Urizar et al., 2011 (link); Yano et al., 2018 (link)). Briefly, cells were prepared in 96-well plates. 5 μM coelenterazine h was added to each well. Three minutes after addition of coelenterazine h (Nanolight), ligands [(+)-pentazocine (Sigma), PD 144418 (Tocris), and haloperidol (Tocris)] were added to each well in serial dilution. BRET was measured as a ratio between measurements at 535 nm for fluorescence (gain 2500, interval time 0.9) and at 485 nm for luminescence (gain 2500, interval time 0.9) using a PHERastar FSX reader (BMG). Results are calculated for the BRET change (BRET ratio for the corresponding drug minus BRET ratio in the absence of the drug). Emax values are expressed as the basal subtracted BRET change in the dose-response graphs.
3
Pharmacological Modulation of Behavioral Outcomes
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PCP (10 mg/kg, subcutaneously; Tocris, Bristol, UK), haloperidol (0.1 mg/kg, intraperitoneally (i.p.); Tocris) and clozapine (10 mg/kg, i.p.; Tocris) were dissolved in physiological saline (NaCl 0.9%) and administered for 5 consecutive days. At the end of treatment (2–3 h after the final drug administration) animals were used for behavioural assessments and biochemical experiments.
4
Signaling Pathway Antibodies and Inhibitors
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PREX1 [D8O8D] rabbit monoclonal, PTEN [138G6] rabbit monoclonal, Phospho-PAK1 (Ser144)/PAK2 (Ser141) rabbit polyclonal, PAK2 [3B5] mouse monoclonal were purchased from Cell Signaling (Danvers, MA, USA). GAPDH [6C5] mouse monoclonal and CDC42 [M152] mouse monoclonal were purchased from Abcam (Cambridge, MA, USA). Phospho-PKCι (T555)/PKCλ (T563) rabbit polyclonal was purchased from Invitrogen (Carlsbad, CA, USA) and PKCι mouse monoclonal from (BD Transduction Laboratories (Mississauga, ON, Canada). Sox2 mouse monoclonal was from R&D Systems (Minneapolis MN, USA). Rac1[23A8] mouse monoclonal, anti-Flag M2 mouse monoclonal and Stem121 mouse monoclonal were purchased from Millipore (Temecula, CA, USA), Sigma-Aldrich (Oakville, ON, Canada) and StemCells Inc. (Newark, CA, USA), respectively. Human brain cerebral cortex protein medley was purchased from Clontech (Mountain View, CA, USA). The following inhibitors were used in the study: gallein (Santa Cruz Biotechnology, CA, USA), BKM120 (Sigma-Aldrich, Oakville, ON, Canada); trichostatin A (Cayman Chemical Company, Ann Arbor, MI, USA); haloperidol, L-741,626 and L-745,870 (Tocris Bioscience, Minneapolis, MN, USA).
5
Pharmacological Modulation of Neurotransmitter Receptors
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All injections were performed intraperitoneally (ip) with a volume of 10 ml/kg body weight. The experimenter was not blind to group-drug allocation. Solutions were freshly prepared on the experimental day. Clozapine-N-oxide (CNO, 1 mg/kg, Tocris, #4936) was dissolved in sterile physiological saline. The preferential D2 antagonist haloperidol (“halo”, 0.5 mg/kg, Tocris, #0931) was dissolved in 5 µl of 0.5% lactic acid and saline. The D1 antagonist SCH23390 (“SCH” [R( + )-7chloro-8-hydroxy-3-methyl-1-phenyl- 2,3,4,5-tetrahydro-1H-3-benzazepine], 0.25 mg/kg, Tocris, #0925) was dissolved in saline. Control groups were injected with either CNO or saline (with or without lactic acid, accordingly).
6
Behavioral Pharmacology: Neuroleptic Drugs and 4-OH-Tamoxifen
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Clozapine (catalog number 0444) stock: 50 mg/ml, Haloperidol (catalog number 0931) stock: 10 mg/ml, and Olanzapine (catalog number 4349) stock: 50 mg/ml were from Tocris Bioscience and dissolved in DMSO. Loxapine (catalog number L106) stock: 10 mg/ml was from Sigma-Aldrich and dissolved in 0.9% saline. All aqueous solutions were buffered to pH 6–6.5 if required. The drugs were administered intraperitoneally.
4-OH-Tamoxifen (catalog number H6278) was purchased from Sigma-Aldrich. It was prepared as described previously (Guenthner et al., 2013 (link)). Briefly, 4-OH-Tamoxifen was dissolved in ethanol at 20 mg/ml and stored at -20°C. 4-OH-Tamoxifen stock in ethanol was mixed with corn oil to achieve the final concentration of 10 mg/ml. The ethanol was evaporated using Centrivap before injection.
4-OH-Tamoxifen (catalog number H6278) was purchased from Sigma-Aldrich. It was prepared as described previously (Guenthner et al., 2013 (link)). Briefly, 4-OH-Tamoxifen was dissolved in ethanol at 20 mg/ml and stored at -20°C. 4-OH-Tamoxifen stock in ethanol was mixed with corn oil to achieve the final concentration of 10 mg/ml. The ethanol was evaporated using Centrivap before injection.
7
Pharmacological Manipulation of Rodent Behavior
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(5R,10S)-(+)-5-Methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate ((+)-MK801, 0.3 mg/kg) and haloperidol (0.5 mg/kg) were purchased from Tocris Bioscience, and D-amphetamine hemisulfate salt (2.5 mg/kg), clozapine N-oxide (CNO, 1 mg/kg), and tamoxifen (100 mg/kg) from Sigma-Aldrich. All drugs were administered intraperitoneally in a volume of 10 ml/kg and dissolved in 0.9% (w/v) NaCl (saline), except CNO that was dissolved in 0.1% DMSO and tamoxifen that was dissolved in sunflower oil/ethanol (10:1) to a final concentration of 10 mg/ml.
8
Haloperidol-Induced Catalepsy in Rodents
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The D2 antagonist haloperidol (Tocris Bioscience, Bristol, United Kingdom) was used to induce cataleptic states. haloperidol was dissolved in physiological saline (0.9%) containing 2% Tween 80 to obtain concentrations of 1 and 2 mg/ml. Injections were administered intraperitoneally in a constant volume of 1 ml/kg, 15 min before the beginning of the tests. The control group received an equivalent volume of physiological saline containing 2% Tween 80 (vehicle). The drug doses were based on previous studies (Colombo et al., 2013 (link); Barroca et al., 2019 (link); Waku et al., 2021 (link)).
9
Neurotransmitter Receptor Binding Assay
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Compounds used in this study were purchased from Sigma-Aldrich (St. Louis, MO) unless indicated otherwise: HEPES, sodium pyruvate (Gibco/ThermoFisher Scientific, Pittsburgh, PA), penicillin/streptomycin, 2-mercaptoethanol, D-glucose, bovine serum albumin (BSA; Merck Millipore, Darmstadt, Germany), EDTA, ascorbic acid, S-(−)-propranolol, R-(−)-deprenyl hydrochloride, pargyline hydrochloride, clorgyline hydrochloride, yohimbine (Tocris, Bristol, United Kingdom), haloperidol, clozapine, olanzapine (Tocris), and [3H]RX821002 (Perkin Elmer, Billerica, MA).
10
Measuring BRET in Receptor Binding Assays
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Acceptor saturating BRET is performed as described previously [22] (link). Briefly, cells were distributed in 96-well plates. Expression of venus fusion proteins was estimated by measuring fluorescence at 535 nm with excitation at 485 nm. Expression of NL fusion proteins was estimated by measuring the luminescence of the cells after incubation with 5 µM coelenterazine h (Nanolight) for 30 min. In parallel, BRET was measured as a ratio between measurements at 535 nm for fluorescence and at 485 nm for luminescence using a Pherastar FSX reader (BMG). Results were plotted as fluorescence over luminescence vs. basal-subtracted BRET ratio.
Drug-induced BRET was conducted as reported previously [26] (link). Briefly, cells were prepared in 96-well plates as in acceptor-saturating BRET. 5 µM coelenterazine h was added to each well. Three minutes after addition of coelenterazine h, ligands [(+)-pentazocine (Sigma), PD144418 (Tocris), and haloperidol (Tocris)] in series of dilution were added to each well. BRET was measured as in acceptor-saturating BRET after 30 min incubation. Results were calculated for the BRET change (BRET ratio for the corresponding drug minus BRET ratio in the absence of the drug). Emax values are expressed as the basal subtracted BRET change in the dose-response graphs.
Drug-induced BRET was conducted as reported previously [26] (link). Briefly, cells were prepared in 96-well plates as in acceptor-saturating BRET. 5 µM coelenterazine h was added to each well. Three minutes after addition of coelenterazine h, ligands [(+)-pentazocine (Sigma), PD144418 (Tocris), and haloperidol (Tocris)] in series of dilution were added to each well. BRET was measured as in acceptor-saturating BRET after 30 min incubation. Results were calculated for the BRET change (BRET ratio for the corresponding drug minus BRET ratio in the absence of the drug). Emax values are expressed as the basal subtracted BRET change in the dose-response graphs.
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