In gramicidin perforated experiments and in experiments performed for assessing the KCC2 efficacy, GABAAR responses were isolated by using a cocktail of drugs containing 0.2 μM tetrodotoxin (TTX, Latoxan Laboratory, France), 4 mM kynurenic acid (Millipore Sigma), 10 μM (+)-tubocurarine (Millipore Sigma), 5 μM Dihydro-β-erythroidine hydrobromide (DHβE, Bio-techne, France), and 3 μm strychnine (Millipore Sigma) that respectively blocked voltage-dependent Na+ action potentials, and glutamate, cholinergic, and glycinergic input to MNs. In CsCl experiments, IPSCs were isolated pharmacologically using DL-AP5 40 μM ((2R)-amino-5-phosphonovaleric acid, Bio-techne, France) and CNQX 20 μM (6-cyano-7-nitroquinoxaline-2,3-dione, Bio-techne, France). mIPSCs were isolated in the presence of 0.2 μM TTX (Latoxan, France). GABA and glycine mIPSCs were isolated by adding 3 μM strychnine or 3 μM GABAzine (SR 95531 hydrobromide, Bio-techne, France), respectively. These blockers were added to the cocktail containing 0.2 µM TTX, 4 mM kynurenic acid, 10 μM (+)-tubocurarine and 5 μM DHβE. In experiments assessing the KCC2 efficacy, 10 µM bumetanide (Millipore Sigma) was applied to block NKCC1 and 10 µM VU0240551 (Bio-techne, France) (10 µM) to block KCC2. Serotonin (5-HT, 10 µM), dopamine (DA, 100 µM) and N-Methyl-D-aspartic acid (NMDA, 10 µM) were from Millipore Sigma.
N methyl d aspartic acid
Manufactured by Merck Group
Sourced in United States
N-methyl-D-aspartic acid is a synthetic amino acid compound commonly used in laboratory research settings. It acts as an agonist for the N-methyl-D-aspartate (NMDA) receptor, which is involved in various neurological processes. This product can be utilized for experimental purposes within the guidelines and protocols established for its intended use in research applications.
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Lab products found in correlation
Mk 801, by Merck Group (4 mentions)
Bicuculline, by Merck Group (3 mentions)
Fluoxetine, by Merck Group (3 mentions)
Picrotoxin, by Merck Group (2 mentions)
Glutamate, by Merck Group (2 mentions)
Forskolin, by Merck Group (2 mentions)
Recombinant mouse leptin, by R&D Systems (2 mentions)
L glutamic acid monosodium salt hydrate, by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Ramelteon, by Merck Group (2 mentions)
Ondansetron, by Merck Group (2 mentions)
Agomelatine, by Merck Group (2 mentions)
Reboxetine, by Merck Group (2 mentions)
Haloperidol, by Merck Group (2 mentions)
Prazosin, by Merck Group (2 mentions)
Im hydrochloride, by Merck Group (2 mentions)
Para chlorophenylalanine, by Merck Group (2 mentions)
Dihydro β erythroidine hydrobromide dhβe, by Bio-Techne (1 mentions)
Vu0240551, by Bio-Techne (1 mentions)
22 protocols using n methyl d aspartic acid
1
Isolating GABAAR Responses for Functional Analysis
2
Intravitreal Injection and Optic Nerve Crush in Mice
3
Pharmacological Manipulation of Neuronal Activity
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Phenylmethanesulfonyl fluoride, aprotinin, phosphatase inhibitor cocktails, CNQX, (2R)-amino-5-phosphonovaleric acid (AP5), N-methyl-D-aspartic acid (NMDA), glutamate, picrotoxin, bicuculline, Forskolin, and Rolipram were from Sigma-Aldrich (St. Louis, USA). Tetrodotoxin (TTX) was from Tocris Bioscience (Ellisville, USA). Detailed information on the antibodies used is provided in the Supplementary Methods.
4
Fluoxetine and Yueju Pill Effects
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Fluoxetine (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 0.9% saline. The OCT Yueju pills (Jiangsu 707 Natural Pharmaceutical Co. Ltd., lot number 150801) were ground into powder and dissolved with 0.9% saline with different doses. The solutions of the Yueju, Fluoxetine, and vehicle were administered to the mice via intragastric administration, and the concentration of the solution was 1 mg/ml. The dose for Fluoxetine was 18 mg/kg/day, and N-Methyl-D-aspartic acid (Sigma-Aldrich, St. Louis, MO, USA) was administered by intracerebroventricular infusion (i.c.v.), 1 pmol/3 μl. All drugs were dissolved in 0.9% saline.
5
Signaling Pathway Modulators Protocol
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5-Bromo-2′-deoxyuridine (BrdU), (±)-Epinephrine hydrochloride, A23187, glutamate, forskolin, isobutyl-1-methylxanthine (IBMX), 4-(3-Butoxy-4-methoxybenzyl) imidazolidin-2-one (Ro 20-1724) (±)-ketamine hydrochloride and N-Methyl-D-aspartic acid (NMDA) were purchased from Sigma–Aldrich. Triethylammonium salt (cAMPS-Rp) was obtained from Tocris Bioscience, and Quest Fluo-8TM AM from AAT Bioquest. All compounds were dissolved in dimethyl sulfoxide (DMSO; Carl Roth). Recombinant human IGF2 protein was purchased from R&D Systems and reconstituted in phosphate-buffered saline (PBS).
6
NMDA Dosage and Administration
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N-methyl-D-aspartic acid (NMDA) was purchased from Sigma-Aldrich Co (St. Louis, MO, USA). The drug was dissolved in physiological saline (0.9% w/v NaCl) at varying doses (50 or 75 mg/kg) and either saline or NMDA was administered via the intraperitoneal (i.p.) route.
7
Intraperitoneal Leptin and Hippocampal Injection Protocol
8
Intraperitoneal Leptin and Hippocampal Injection Protocol
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Recombinant mouse leptin (R&D Systems, Minneapolis, MN) was dissolved in sterile saline at a concentration of 1.0 mg/ml and administered intraperitoneally (i.p.) at a dose of 1.0 mg/kg body weight. For intra-CA3 injections, N-methyl-D-aspartic acid (NMDA) and l5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801; Sigma-Aldrich, St Louis, MO) were dissolved in Dulbecco's phosphate-buffered saline and injected at a dose of 0.1 nmol/μl for NMDA and a dose of 1 μg/μl for MK-801. For intra-DG injection, Akt inhibitor VIII (1,3-dihydro-1-(1-((4-(6-phenyl-1H-imidazo[4,5-g]qui-noxalin-7-yl)phenyl)methyl)-4-piperidinyl)-2H-benzimidazol-2-one), an isozyme-selective Akt inhibitor specific for Akt1/2 (Calbiochem, La Jolla, CA), was dissolved in DMSO (Sigma) and injected at a dose of 3.0 μg/μl. L-glutamic acid monosodium salt hydrate and ascorbic acid were purchased from Sigma and dissolved in distilled water in stock concentrations of 5 mM and 100 mM for biosensor calibration, respectively.
9
Ebselen Modulates NMDA-Induced Osteoclast Formation
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Ebselen (2-phenyl-1,2-benzisoselenazol-3[2H]-one) was purchased from Cayman Chemical (Ann Arbor, MI, USA) with high purity (more than 99%) and was dissolved in dimethyl sulfoxide. N-Methyl-D-aspartic acid (NMDA) receptor agonist, NMDA was purchased from Sigma (St. Louis, MO, USA) with high purity (more than 98%). Recombinant soluble human macrophage colony-stimulating factor (M-CSF), RANKL, and IL-1α were obtained from PeproTech EC Ltd. (London, UK). A monoclonal antibody against β-actin was obtained from Sigma (St. Louis, MO, USA). Antibodies against phosphorylated (phospho)-p38, p38, phospho-extracellular signal-regulated protein kinase (ERK), ERK, phospho-JNK, JNK, phospho-IκB, phospho-phosphoinositide 3-kinase (PI3K), PI3K, phospho-Akt, Akt, caspase-3, and caspase-9 were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Antibodies against c-Fos, nuclear factor of activated T cells c 1 (NFATc1), and IκB were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Fetal bovine serum (FBS), α-minimum essential medium (α-MEM), penicillin, and streptomycin were purchased from Gibco BRL (Grand Island, NY, USA). All other chemicals were of analytical grade or complied with the standards required for cell culture experiments.
10
Neuronal Responses to Pharmacological Agents
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GM was purchased from Standard Technology Co., Ltd (Shanghai, China). The purity of the synthetic GM was ≥98%, as assessed by HPLC. The compound was dissolved and stored in dimethyl sulfoxide (DMSO) to produce 20 mmol/L stock solutions that were then diluted in bath solution to obtain the final concentrations. DMSO at the final concentrations (≤0.5%) was well tolerated with no observable toxic effects to the neurons. To conduct research on animals, GM was dissolved in a mixture of 5% DMSO and 95% Tween-80. TTX was purchased from Aquaculture Technical Developing Company, Qinhuangdao, China. Topiramate (TPM), levetiracetam (LEV), γ-amino butyric acid (GABA), N-methyl-D-aspartic acid (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), kainic acid (KA) and acetylcholine chloride (ACh-Cl) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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