(S)-(+)-α-Amino-4-carboxy-2-methylbenzeneacetic acid (LY-367385), 2-Methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP), D-(−)-2-Amino-5-phosphonopentanoic acid (D-AP5), 3,5-dihydroxyphenylglycine (RS-3,5-DHPG), 8,8’-[Carbonylbis[imino-3,1-phenylenecarbonylimino(4-methyl-3,1-phenylene)carbonylimino]]bis-1,3,5-naphthalenetrisulfonic acid hexasodium salt (suramin), and Protease-Activated Receptors TFLLR-NH2 were purchased from Tocris. The Ca2+ indicator Fluo-4-AM was purchased from Life Technologies Ltd. Picrotoxin, atropine, ATP and other drugs were purchased from Sigma-Aldrich.
Suramin
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
Suramin is a lab equipment product manufactured by Bio-Techne. It is a synthetic polysulfonated naphthylurea compound that functions as a competitive antagonist of various growth factors and enzymes. Suramin can inhibit the activities of certain proteins involved in cellular processes. The core function of Suramin is to act as a research tool for studying the effects of growth factor and enzyme inhibition in in vitro and in vivo experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Arl67156, by Bio-Techne (5 mentions)
Ppads, by Bio-Techne (5 mentions)
Fluo 4 am, by Thermo Fisher Scientific (4 mentions)
5 bdbd, by Bio-Techne (3 mentions)
Mrs2500, by Bio-Techne (3 mentions)
Adenosine triphosphate (atp), by Merck Group (2 mentions)
Carbenoxolone, by Merck Group (2 mentions)
Apyrase, by Merck Group (2 mentions)
Cgs15943, by Bio-Techne (2 mentions)
A438079, by Bio-Techne (2 mentions)
Atpγs, by Bio-Techne (2 mentions)
Ryanodine, by Bio-Techne (2 mentions)
Nf157, by Bio-Techne (2 mentions)
Ar c 118925xx, by Bio-Techne (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Picrotoxin, by Merck Group (1 mentions)
Tfllr nh2, by Bio-Techne (1 mentions)
Fast green dye, by Merck Group (1 mentions)
20 protocols using suramin
1
Pharmacological Modulation of Neuronal Signaling
2
In Utero Calcium Imaging of Mouse Embryos
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Pregnant C57bl/6J mice at the 13–15th day of gestation (E13–E15) were anesthetized with intraperitoneal injection of ketamine/xylazine (80/10 mg/kg) and placed on a heating pad at 37°C.
A single 15–20 mm incision was made in the abdomen, the uterine horn exposed and embryos injected ventricularly with Fluo-4AM in loading solution using a capillary glass pipette. The loading solution consisted of 17 μg of calcium dye Fluo-4AM (Molecular Probes) dissolved in 3 μl of 20% F-127 pluronic® acid (Sigma) in dimethyl sulfoxide (DMSO; Sigma) that was then diluted in artificial cerebrospinal fluid (ACSF; in mM: 125 NaCl, 1.25 NaH2PO4, 2 CaCl2, 1 MgCl2, 5 KCl, 20 D-glucose, 10 HEPES) to reach a final concentration 0.4 mM of Fluo-4. 13 mM FastGreen dye (Sigma) was added to the solution for visual injection guidance. Injections were performed with a Picospritzer II, 1–2 μl per embryo depending on the size of the embryo through a glass pipette with a 12 mm long pulled tip that measured ~50 μm in tip diameter (Drummond Scientific) targeting the lateral ventricle. The uterine horn was rinsed with 37°C ACSF every 2 min to prevent hypothermia. After all embryos had been injected with calcium dye the uterine horn was placed back inside the mother and the cavity was sutured. In some experiments suramin (3.7 mM, Tocris Bioscience) was added to the injected solution.
A single 15–20 mm incision was made in the abdomen, the uterine horn exposed and embryos injected ventricularly with Fluo-4AM in loading solution using a capillary glass pipette. The loading solution consisted of 17 μg of calcium dye Fluo-4AM (Molecular Probes) dissolved in 3 μl of 20% F-127 pluronic® acid (Sigma) in dimethyl sulfoxide (DMSO; Sigma) that was then diluted in artificial cerebrospinal fluid (ACSF; in mM: 125 NaCl, 1.25 NaH2PO4, 2 CaCl2, 1 MgCl2, 5 KCl, 20 D-glucose, 10 HEPES) to reach a final concentration 0.4 mM of Fluo-4. 13 mM FastGreen dye (Sigma) was added to the solution for visual injection guidance. Injections were performed with a Picospritzer II, 1–2 μl per embryo depending on the size of the embryo through a glass pipette with a 12 mm long pulled tip that measured ~50 μm in tip diameter (Drummond Scientific) targeting the lateral ventricle. The uterine horn was rinsed with 37°C ACSF every 2 min to prevent hypothermia. After all embryos had been injected with calcium dye the uterine horn was placed back inside the mother and the cavity was sutured. In some experiments suramin (3.7 mM, Tocris Bioscience) was added to the injected solution.
3
Dissecting Metabolic Pathways through Pharmacological Modulation
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CL316243, forskolin, 8Br-cAMP, ARL 67156 trisodium salt, and suramin were procured from Tocris (Minneapolis, MN). Carbenoxolone, trovofloxacin, and spironolactone were from Sigma Aldrich (St. Louis, MO). Silencer select siRNAs against mouse Panx1 (assay ID-s79985) or mouse Gβ subunits (GNB1 – assay ID –ss66813; GNB2 – assay ID –ss66816; GNB3 – assay ID –ss66822; GNB4 – assay ID –n420113) and silencer select negative control-2 siRNA (4390846) were purchased from ThermoFisher Scientific (Middletown, VA). Antibodies used in the study were as follows: mouse monoclonal anti-Panx1 (Clone 720505, #MAB7097) was from R&D Systems (Minneapolis, MN); rabbit-polyclonal anti-UCP1 (#U6382) was from Sigma–Aldrich (St. Louis, MO); mouse monoclonal OXPHOS antibody cocktail (#MS604) was from Mitosciences (Eugene, OR); rabbit monoclonal anti-Panx1 antibody (Clone D9M1C, #91137), anti-PKC phosphorylation antibody sampler kit (#9921) and rabbit monoclonal anti-vinculin (Clone E1E9 V, #13901) were from Cell Signaling (Danver, MA). Cellular glucose uptake was measured using a glucose uptake-Glo assay kit (#J1341, Promega, Madison, WI).
4
Murine Dendritic Cell Characterization
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Female C57BL/6 mice purchased from Ji'nan Pengyue Laboratory Animal Breeding Co., Ltd. (China) were housed and maintained in the animal facilities of Weifang Medical University. Recombinant murine GM-CSF and IL-4 were purchased from R&D Systems (USA). A Fluo-4 Direct™ calcium assay kit was purchased from ThermoFisher Scientific (USA). ARL-67156, Bz-ATP, OX-ATP, and suramin were purchased from Tocris Bioscience (USA). A-438079, a competitive P2X7 receptor antagonist, was purchased from Santa Cruz (USA). Luminescent ATP Detection Assay kit was purchased from Abcam (USA). The endocytosis inhibitors MDC, PitS2, Dynasore, and P2X4-specific antagonist PSB-12062 were purchased from Sigma-Aldrich (USA). RNAiso Plus, SYBR Premix Ex Taq II, and the PrimeScriptTM RT reagent Kit were obtained from Takara (China). CD11c-APC (#550261), CD11b-FITC (#553310), MHC II-PE (#558593), and F4/80-PE (#565410) and isotype control antibodies were purchased from BD Bioscience (USA). CD39-PE (#12-0391-82) was purchased from Thermo Fisher (USA). All experiments involving animals were performed in accordance with the Chinese National Laboratory Animal-Guideline for Ethical Review of Animal Welfare and approved by the Institutional Animal Care and Use Committee of Weifang Medical University (NO. 010/2017).
5
Standardized Cell Culture Reagents
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Standard chemicals and cell culture reagents were purchased from Sigma and ThermoFisher. ARL67156, suramin, and PPADS were purchased from Tocris.
6
Microglial Cell Migration Assay
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Migration assays were performed in the Boyden 10 well chemotaxis chamber (Neuro Probe, Gaithersburg, MD, USA), according to the manufacturer’s instructions. Briefly, complete DMEM medium (supplemented with EV-depleted 10% FBS and 1% p/S) was added to the lower wells and separated from the upper wells by a polycarbonate filter membrane with 8 µm size pores. Microglial cells were plated into the upper wells (30,000 cells in 285 μL of complete medium per well), left overnight, and then pre-treated for 30 min with one of the following inhibitors: 200 µM suramin, 1 µM AR-C66931, 10 µM 5-BDBD (all from Tocris, Bristol, UK), or with 10 µM of cilengitide (Sigma, Burlington, MA, USA) for 2 h. Afterwards, cells were treated with 1 AU of EVs and then left overnight. For the apyrase treatments, 20 U/mL apyrase (Sigma) and 1 AU of EVs were added simultaneously to the upper wells and cells incubated overnight. After incubation, membranes were washed 3 times with PBS, fixed with methanol for 5 min, air-dried, stained with 0.25% Cristal Violet for 15 min, and again washed 3 times with PBS. The number of cells that migrated to the opposite side of the membrane was determined using an optical microscope in three randomly chosen fields from at least three wells for each experimental group.
7
Immortalized Human Keratinocyte Culture
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Keratinocytes (courtesy of Leena Bruckner-Tuderman, Department of Dermatology, University Hospital Freiburg) isolated from human foreskin were transformed with the human papillomavirus oncogenes E6 and E7 for immortalization (Kaur and McDougall, 1988 (link)). Cells were kept in Keratinocyte Serum-Free Medium (KSFM; Invitrogen, Carlsbad, CA) supplemented with 0.3 ng/ml recombinant EGF and 30 ng/ml bovine pituitary extract and 100 U/ml penicillin/streptomycin (culture medium). The experiment medium contained an additional 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Sigma-Aldrich, St. Louis, MO). For pH experiments, the medium was adjusted to a pH of 7.45 by 2.5 N NaOH or to 6.6 by 10% HCl. The inhibitors CK-666 (Sigma-Aldrich), NSC-23766 (Tocris, Bristol, UK), ZCL-278 (Tocris), PIK-90 (Selleck Chemicals, Houston, TX), AG-1478 (Cayman Chemicals, Ann Arbor, MI), Y-27632 (Merck-Millipore, Billerica, MA), (–)-blebbistatin (Sigma-Aldrich), suramin (Tocris), pertussis toxin (Enzo Scientific, Farmingdale, NY), gallein (Tocris), telenzepine (Tocris), PPADS (Tocris), and CK-689 (Merck-Millipore) were used at the indicated concentrations.
8
Standard and High-KCl Extracellular Solutions
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Standard ECS consisted of the followings: 136 mM NaCl, 5 mM KCl, 2.5 mM CaCl2, 0.5 mM MgCl2, 1.2 mM NaH2PO4, 11 mM glucose, and 12 mM NaHCO3 (328 mOsm/L). To obtain high-KCl solutions, we increased the extracellular K+ in the standard ECS to 100 mM and reduced extracellular Na+ in to 41 mM. The Hanks’ balanced salt solution consisted of the following ingredients: 137 mM NaCl, 5.0 mM KCl, 2.0 mM CaCl2, 0.5 mM MgCl2, 0.44 mM KH2PO4, 0.34 mM Na2HPO4, 4.17 mM NaHCO3, and 5.55 mM glucose. The pH of these solutions was adjusted to 7.4 using Tris (Wako Pure Chemicals, Osaka, Japan). Suramin and MK801 were obtained from Tocris Bioscience (Ellisville, MO, United States). Stock solutions for these agents were prepared in dimethyl sulfoxide or Milli-Q water (Merck KGaA, Darmstadt, Germany), and were later diluted to the appropriate concentrations in either ECS. Except where indicated, all reagents were obtained from Sigma Chemical Co., (St. Louis, MO, United States).
9
Comprehensive Calcium Signaling Assay
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Apamin, cyclopiazonic acid (CPA), GSK2193874, GSK1016790A, HC067047, NS309, 1400W, Nω‐propyl‐l ‐arginine hydrochloride, ODQ, RN1747, Rp‐8‐Br‐PET‐cGMPS, Suramin, and Tram 34 were purchased from Tocris Bioscience (Minneapolis, MN). ATP and Nω‐nitro‐l ‐arginine were obtained from Sigma‐Aldrich (St. Louis, MO). DAF‐FM diacetate, Fluo‐4AM (Ca2+ indicator), and EGTA‐AM (Ca2+ chelator) were purchased from Fischer Scientific (Pittsburgh, PA). Spermine NONOate and U46619 were purchased from Cayman Chemical (Ann Arbor, MI). All other chemicals were obtained from Sigma‐Aldrich (St. Louis, MO).
10
Reagents for Biological Experiments
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Collagenase P and FA-free Bovine serum albumin were obtained from Roche Diagnostics S.L. (Barcelona, Spain). Bovine serum albumin and most of the substances, inhibitors (POM–1, NPPB, carbenoxolone, flufenamic acid, mefloquine, diazoxide), activators (bzATP), enzymes and coenzymes were obtained from Sigma-Aldrich Química S.A. (Madrid, Spain). Other inhibitors used (ARL–67156, CGS15943, suramin) were from Tocris Bioscience (Biogen Científica S.L., Spain). Rat insulin standards were from Linco Research, Inc. (St. Charles, Missouri, U.S.A.). Na125I was obtained from PerkinElmer España, S.L. (Madrid, Spain). Ad-CMV-Luciferase was from Vector Biolabs (Philadelphia, PA, U.S.A.). Inorganic compounds and organic solvents were obtained from VWR International Eurolab S.L. (Spain).
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