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).
L 745870
Manufactured by Bio-Techne
Sourced in United States, Switzerland, United Kingdom
L-745870 is a selective dopamine D4 receptor antagonist. It has a high affinity for the D4 receptor and low affinity for other dopamine receptor subtypes.
Automatically generated - may contain errors
Lab products found in correlation
Sulpiride, by Bio-Techne (4 mentions)
Quinpirole, by Bio-Techne (3 mentions)
L 741 626, by Bio-Techne (2 mentions)
Sch23390, by Bio-Techne (2 mentions)
Tamoxetine, by Bio-Techne (2 mentions)
Gbr12783, by Bio-Techne (2 mentions)
Way 100635, by Abcam (2 mentions)
Citalopram, by Bio-Techne (2 mentions)
Sumatriptan, by Bio-Techne (2 mentions)
Gabazine, by Abcam (2 mentions)
Kynurenic acid, by Abcam (2 mentions)
Bkm120, by Merck Group (1 mentions)
Haloperidol, by Bio-Techne (1 mentions)
Human brain cerebral cortex protein medley, by Takara Bio (1 mentions)
Trichostatin a, by Cayman Chemical (1 mentions)
Gallein, by Santa Cruz Biotechnology (1 mentions)
Pramipexole, by Bio-Techne (1 mentions)
Skf81297, by Bio-Techne (1 mentions)
Pd 168077, by Bio-Techne (1 mentions)
D ap5, by Bio-Techne (1 mentions)
10 protocols using l 745870
1
Signaling Pathway Antibodies and Inhibitors
2
Dopamine Receptor Agonist and Antagonist Effects
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The DR1 agonist SKF-81297 (1 μM), the D2-like receptor agonist quinpirole (10 μM), the selective DR3 agonist pramipexole (10 μM), the DR4 agonist PD-168077 (200 nM), the selective DR4 antagonist L-745870 (500 nM), the dopamine (200 μM), the n-methy-d-aspartate (NMDA) receptor antagonist D-AP5 (50 μM) and CCH (5 μM) were purchased from Tocris Bioscience. Stock solutions, at thousand times the final concentration, were made up in water or in DMSO, and stored in individual aliquots at 20°C. Final solutions were prepared freshly on the day of the experiment. Drugs were applied to the recording ACSF 15 min after steady-state γ power was reached. The dead volume and bath volume was kept such that final concentration was reached in the bath in about 5 min.
3
Insulin Receptor Antibody Immunoblotting
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Insulin receptor antibody is polyconal rabbit antihuman antipeptide (Santa Cruz Biotechnology, Santa Cruz, CA).18 Goat polyclonal p‐insulin Rβ antibody (Santa Cruz Biotechnology) was used as the phosphorylated insulin receptor antibody. RPT cells were treated with vehicle (dH2O), a D4 receptor agonist (PD168077; Tocris Cookson Ltd, Bristol, UK),19 or ABT724 (2‐[(4‐pyridin‐2‐ylpiperazin‐1‐yl)methyl]‐1H‐benzimidazole trihydrochloride; Sigma‐Aldrich, St. Louis, MO) or a D4 receptor antagonist20 (L‐745870 [Tocris] or L750667 [Sigma‐Aldrich]) at the indicated concentrations and times as described previously.21 Transblots were probed with insulin receptor antibody (1:400). The amount of protein transferred onto membranes was determined by immunoblotting for α‐actin.
4
Pharmacological Modulation of Neurotransmission
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Carbachol (carbamylcholine chloride), kainate (kainic acid monohydrate), quinpirole, (+)- sulpiride, (+)-SKF-38393, PD168,077, carbenoxolone disodium salt, picrotoxin were purchased from Sigma (Sigma-Aldrich, Buchs, Switzerland); SYM2206 and L745.870 were from Tocris Bioscience (Bristol, UK); AP-5 (DL-2-amino-5-phosphopentanoic acid) was from Alexis Biochemicals (San Diego, CA); R(+)-SCH23390 HCl was from Research Biochemical International (Sigma-Aldrich, Buchs, Switzerland).
5
Retinal Dopamine Receptor Pharmacology
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(–)-Sulpiride, (–)-eticlopride, and L-745,870 were purchased from Tocris Bioscience (Minneapolis, MN). Eticlopride and L-745,870 were dissolved in physiological saline (0.9% NaCl). Sulpiride was dissolved in dimethylsulphoxide (DMSO) and diluted 50-fold in physiological saline. The drug solutions were applied to the bathing solution using calibrated syringe pumps (Razel Scientific Instruments). The final concentration of DMSO at the retina for the sulpiride solution was approximately 0.02%. Drugs were bath applied for ~10 min to ensure stable responses before effects were examined. Only one cell was studied in each retina to avoid possible residual drug effects.
6
Monitoring Dopamine and Glutamate Dynamics in Mice
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Experiments were performed 36 hours after probe implantation in freely moving mice. Ringer’s solution (147 mM NaCl, 4 mM KCl, and 2.2 mM CaCl2) was pumped through the probes at a constant rate of 1 μl/min. After 30 min of equilibration, samples were collected every 10 min (DA analysis) or 20 min (GLU analysis) for 80 min. The indicated doses of (+)-METH-HCl (NIDA Pharmacy) or (−)-cocaine-HCl (NIDA Pharmacy) were administered intraperitoneally using saline (10 ml/kg) (0.9% NaCl) as a vehicle. After the systemic administration, samples were collected every 10 min (DA analysis) or 20 min (GLU analysis) for 90 or 120 min, respectively. Where indicated, vehicle (saline), the D1 receptor antagonist SCH23390 (1 mg/kg; Tocris), or the D4 receptor antagonist L-745,870 (1 mg/kg; Tocris) was systemically administered (10 ml/kg, intraperitoneally) 10 min before METH or cocaine treatment. DA content in the dialyzate fractions was measured by high-performance liquid chromatography (HPLC) coupled to a coulometric detector (5200a Coulochem III, ESA). GLU was measured by HPLC coupled to a GLU oxidase enzyme reactor and electrochemical detector (Eicom Corporation).
7
Pharmacological Reagents Inventory
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Cocaine HCl was obtained from the National Institute on Drug Abuse. NBQX, CPP, sulpiride, quinpirole, sumatriptan, GBR12783, tamoxetine, citalopram, 5-HT, and L-745870 were purchased from Tocris. Gabazine, kynurenic acid (sodium salt), and WAY 100635 were obtained from Abcam. All other chemicals were from Sigma.
8
Pharmacological Reagents Inventory
9
Dopamine Receptor Modulation of Motor Activity
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Dopamine hydrochloride (Sigma-Aldrich, Inc., St. Louis, MO) was bath applied in separate experiments at 1 µM, 3 µM, 10 µM, 30 µM, 100 µM, and 300 µM to determine dose-dependent effects on motor activity. The receptor-selective agonists we used included SKF 81297 for D1-like receptors (10–50 µM; Tocris, Minneapolis, MN); quinpirole for D2-like receptors (10–50 µM; Tocris); and the D1/D2 receptor co-agonist SKF 83959 (10–50 µM; Tocris). For dopamine receptor antagonists we used the D1-like antagonist SCH-23390 (10 µM; Tocris); the D2-like antagonists sulpiride (20 µM) and L-741626 (12 µM); the selective D3 receptor antagonist SB 27701A (5 µM; Tocris); the selective D4 receptor antagonist L-745870 (5 µM; Tocris). We also used the α2 adrenergic receptor antagonist, yohimbine (2–4 µM; Tocris). Endogenous dopamine levels were manipulated with the DAT inhibitor GBR-12909 (10 µM; Hello Bio, Princeton, NJ) and the monoamine oxidase A and B inhibitor bifemelane (50 µM; Tocris).
10
Intraventricular L-745870 Dose Effect
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The L-745870 (D4R selective antagonist; Tocris Bioscience, UK) was used. The drug was initially prepared in a stock solution dissolved in 5% dimethyl sulfoxide (DMSO), and then dissolved in aliquots with sterile saline solution (0.9%). In a previous study performed in our laboratory, the intra-PVN administration of 0.1 µg of L-745870 prevented the PD-168,077-induced hyperfagia (Tejas-Juárez et al., 2014 (link)). Based on this evidence, we increased this dose 10-fold and 20-fold more to evaluate the intraventricular effect of the L-745870. The final concentrations infused in each experiment were 1-µg and 2-µg in a volume of 5 µl at a speed of 1 µl/min in the right lateral ventricle, using a high-precision digital syringe (Hamilton Co., Reno, NV). After administration, the microinjector remained 1-minute in the site to avoid backflow. Control animals received a volume of 5 µl of sterile saline 0.9% + 0.05% DMSO.
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