Recombinant human tPA (Actilyse®) was a generous gift from Boehringer Ingelheim (Auckland, New Zealand) or purchased from Biopur (Reinach, Switzerland). For some experiments the excipients in Actilyse® tPA were removed by dialysis against HEPES (Samson et al., 2008b (link)). NMDA, MK-801 and amino-5-phosphonovalerate (APV) were purchased from Sigma Aldrich (Auckland, New Zealand). Nimodipine, bicuculline and 4-AP were purchased from Tocris (MO, USA). 2,7-Bis-(4-aminobenzylidene)-cycloheptan-1-one dihydrochloride (tPA-STOP) was purchased from American Diagnostica (Greenwich, CT, USA). Human α2-antiplasmin was purchased from MyBioSource (San Diego, CA, USA). Receptor-associated protein (RAP) was produced as a glutathione S-transferase fusion protein and purified by glutathione-affinity chromatography. The glutathione S-transferase tag was removed by thrombin cleavage prior to use. The active site mutant human tPA (S478A) was purchased from Molecular Innovations (MI, USA).
Nimodipine
Manufactured by Bio-Techne
Sourced in United States, United Kingdom
Nimodipine is a calcium channel blocker used as a laboratory reagent. It functions by inhibiting the movement of calcium ions across cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
Mk 801, by Bio-Techne (3 mentions)
D ap5, by Bio-Techne (3 mentions)
Bicuculline, by Bio-Techne (2 mentions)
Thapsigargin, by Bio-Techne (2 mentions)
D serine, by Bio-Techne (2 mentions)
Cgp55845, by Bio-Techne (2 mentions)
Brain derived neurotrophic factor (bdnf), by Bio-Techne (2 mentions)
Picrotoxin, by Bio-Techne (2 mentions)
Mibefradil, by Bio-Techne (2 mentions)
Picrotoxin, by Merck Group (2 mentions)
Mk 801, by Merck Group (1 mentions)
Actilyse, by Boehringer Ingelheim (1 mentions)
Dantrolene, by Merck Group (1 mentions)
Fk506, by Merck Group (1 mentions)
Conotoxin gvia, by Alomone (1 mentions)
Cycloheximide (chx), by Bio-Techne (1 mentions)
Okadaic acid, by Santa Cruz Biotechnology (1 mentions)
Calyculin a, by Cell Signaling Technology (1 mentions)
Rapamycin, by Cell Signaling Technology (1 mentions)
2 apb, by Bio-Techne (1 mentions)
24 protocols using nimodipine
1
Recombinant tPA Characterization Protocols
2
Synthetic Aβ Oligomers: A Tractable Tool for Alzheimer's Research
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Full-length, synthetic Aβ 1–42 peptides (AβOs; American Peptide, Sunnyvale, CA) were prepared exactly according to the method of Lambert et al. (1998) (link) and applied to 11–13 DIV cells at a final concentration of 500 nM for 18 h. We use synthetic AβOs in our studies for the following reasons. First, they mimic the toxic properties of natural oligomers (brain or cell derived) as described previously (Jin et al., 2011 (link); Welzel et al., 2014 (link)). Second, unlike natural oligomers, synthetic AβOs can be detected by immunocytochemistry. Confirmation of AβO binding is crucial in our experiments because it varies considerably between neurons. Although they may not be identical to natural oligomers, synthetic AβOs are a tractable tool for investigating mechanisms of AD pathogenesis (Ferreira and Klein, 2011 (link)). After AβO or vehicle exposure, cells were incubated with 1 μM FK506 (Sigma-Aldrich, St. Louis, MO) or equivalent volumes of vehicle (ethanol) for 1–3 h before imaging of transport. For all VGCC inhibition experiments, cells were incubated with 100 μM conotoxin GVIA (Alomone Labs, Jerusalem, Israel), 50 μM agatoxin IVA (Alomone Labs), or 10 μM nimodipine (Tocris Bioscience, Bristol, United Kingdom) for 30 min before AβO treatments. For all RyR inhibition experiments, cells were incubated with 0.5 μM dantrolene (Sigma-Aldrich) for 1 h before AβO treatment.
3
Pharmacological Modulation of Intracellular Pathways
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(RS)-3,5-Dihydroxyphenylglycine (DHPG), NNC 55–0396 dihydrochloride (NNC), nimodipine, 2-aminoethoxydiphenylborane (2-APB) and cycloheximide were obtained from Tocris. Rapamycin and Calyculin-A were from Cell Signaling. Calpain inhibitor III was from Calbiochem. Okadaic acid was from Santa Cruz. All the other reagents were from Sigma. Water insoluble compounds were dissolved in DMSO and diluted to reach a final concentration less than 0.1%.
4
Preparation of Stock Solutions for Ion Channel Experiments
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Stock solutions of Nimodipine (20 mM in DMSO), amiloride (500 mM in DMSO), and cariporide (10 mM in DMSO) were stored at −20 °C, and were diluted at least 1000 times to reach desired final concentrations. Nimodipine and cariporide were purchased from Tocris Cookson (Ellisville, MO, USA), and amiloride from Sigma-Aldrich (St Louis, MO, USA). Na+-free solutions were prepared with total replacement of extracellular Na+ with N-methyl-D-glucamine (NMDG+), and 20 mM K+ solutions were prepared with equal molar substitution of K+ for Na+. All solutions were adjusted to pH 7.4 before use.
5
Pharmacological Modulation of Ion Channels
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20 μM ZD7288 (Abcam) was employed to block HCN channels. 50 μM BaCl2 (Sigma Aldrich) was used to block inward-rectifier potassium channels. 20 μM Riluzole (Abcam) and 10 μM Nimodipine (Tocris Biosciences) were used to block persistent sodium and L-type calcium channels, respectively. For experiments with ZD7288, cells were patched with pipette solution containing 20 μM ZD7288 along with adding it to bath solution (Ashhad and Narayanan, 2016 (link)). For long-term control and TBF experiments in the presence of pharmacological agents, slices were pretreated with the respective pharmacological agent for at least 15 min before the start of recordings.
6
Pharmacological Modulation of Synaptic Plasticity
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Drugs were bath-applied to the whole slice through the perfusion system by dilution of concentrated stock solutions (prepared in water or DMSO) in ACSF, or by adding the drugs to the patch pipette solution when it was applied intracellularly to the postsynaptic cell only. If the drug was not water-soluble, vehicle control experiments were carried out. For each set of recordings, interleaved control and drug conditions were carried out and were pseudorandomly chosen. The following drugs were used in this study: 100 μM dopamine hydrochloride (Sigma–Aldrich, Dorset, United Kingdom), 100 μM D-AP5 (Tocris Bioscience, Bristol, United Kingdom), 10 µM nimodipine (Tocris Bioscience), 1 µM PKA inhibitor fragment (6-22) amide (Tocris Bioscience), 0.5 mM anisomycin (stock solution in EtOH; Tocris Bioscience), and 1 mM MK801 (Tocris Bioscience).
7
Pharmacological Agents for Synaptic Plasticity
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Pharmacological agents were purchased from: Sigma-Aldrich: Fluoroacetate, thapsigargin, BAPTA, bicuculline methbromide, Tricine, Zinc chloride, d -serine, and GDPβS; Tocris Bioscience: nimodipine, (+)-MK-801 maleate, d -AP5, NBQX, TTX citrate, PPDA, Ro 25-6981 maleate, MCPG, MPEP, LY341495, AM251, 2-AG, and FK506; and Abcam: UBP-141. Salts used for internal and external solutions were purchased from Sigma-Aldrich. Compounds were dissolved in H2O or Ringer solution with the exception of thapsigargin, nimodipine, PPDA, FK506, THL, AM251, and 2-AG, which were dissolved in DMSO. Vehicle (DMSO) at the concentrations used did not affect baseline EPSP amplitudes and had no other detectable effects on the neurons. When investigating the effect of pharmacological agents on plasticity, all drugs were included in the superfusion fluid or patch pipette from the start of the experiment until completion (from 0 to 50 min in a standard plasticity experiment), except for Fluoroacetate, which was applied from 60 min before the start of recording. When determining the effect of a pharmacological agent on baseline condition, a stable baseline of at least 10 min was first recorded and then the drug was bath applied by switching to a different perfusion line.
8
Cerebral Vasodilation Therapy for Ischemic Stroke
9
Neurochemical Agents in Neuroscience
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MK-801, NBQX, CGP-55845, nimodipine, DCG-IV, and GYKI 53655 were obtained from Tocris-Cookson. LY 303070 was obtained from ABX advanced biochemical compounds. BoTX was obtained from List Biological. All other chemicals and drugs were purchased from Sigma-Aldrich.
10
Pharmacological Modulation of Neuronal Functions
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Drugs were prepared as stock solutions for frozen aliquots at − 20 °C. They were diluted from the stock solution to the final desired concentration in the ACSF before being applied to the perfusion solution. BDNF (1, 20, 50 ng/ml), nimodipine (10 μM), d (−)-2-amino-5-phosphonopentanoic acid (AP-5, 100 μM), (+)-α-methyl-4-carboxylphenlyglycine (MCPG, 500 μM), K252a (200 nM), NASPM (50 μM), ZIP (5 μM) and NB001 (20 μM) were used in the current study. Drugs were infused during the period of the horizontal bar on the graphs. NB001 was provided by NeoBrain Pharmac Inc (Canada). BDNF, nimodipine, MCPG, K252a, ZIP was purchased from Tocris Bioscience. AP-5, NASPM were purchased from Hello Bio.
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