Ns309

Manufactured by Merck Group

Sourced in United States

NS309 is a lab equipment product manufactured by Merck Group. It is a selective positive modulator of the small conductance calcium-activated potassium (SK) channels.

Automatically generated - may contain errors

Lab products found in correlation

Tram 34, by Merck Group (6 mentions) Adenosine diphosphate (adp), by Merck Group (4 mentions) Apamin, by Merck Group (4 mentions) Endothelin 1, by Merck Group (4 mentions) Substance p, by Merck Group (3 mentions) Potassium chloride (kcl), by Merck Group (3 mentions) Papaverine, by Merck Group (3 mentions) Diltiazem, by MP Biomedicals (3 mentions) Cyppa, by Merck Group (2 mentions) U46619, by Merck Group (2 mentions) Indomethacin, by Merck Group (2 mentions) L name, by Merck Group (2 mentions) Sodium nitroprusside, by Merck Group (1 mentions) Saponin, by Merck Group (1 mentions) Antimycin a, by Merck Group (1 mentions) Sodium ascorbate, by Merck Group (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)

16 protocols using ns309

Pharmacological Modulation of Signaling Pathways

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ADP, substance P, sodium nitroprusside, NS309, TRAM34 and apamin were obtained from Sigma-Aldrich and dissolved in ultrapure distilled water on the day of the study.

Liu Y., Xie A., Singh A.K., Ehsan A., Choudhary G., Dudley S., Sellke F.W, & Feng J. (2015). Inactivation of Endothelial Small/Intermediate Conductance of Calcium-Activated Potassium Channels Contributes to Coronary Arteriolar Dysfunction in Diabetic Patients. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 4(8), e002062.

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Culturing Pancreatic Cancer Cell Lines

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Panc-1 cells were cultured in DMEM with 10% fetal calf serum; AsPC-1 and BxPC-3 cells were cultured in RPMI 1640 with 10% fetal calf serum, Capan-1 cells were cultured in IMDM with 20% fetal calf serum; Mia PaCa-2 cells were cultured in DMEM/F12 with 10% fetal calf serum and 2.5% horse serum; All media used for routine passaging contained GlutaMAX and 25 mM glucose. KCNN4 inhibitor rac-16 was synthesized as described before [26 (link)], NS309 and TRAM-34 were obtained from Sigma-Aldrich (St. Louis, MO, USA), diluted in DMSO to 10 mM stock solutions and stored in -20°C. Oligomycin (Oligo), FCCP (carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone), rotenone (R) and antimycin A (A) were obtained from Sigma-Aldrich (St. Louis, MO, USA), diluted in DMSO to 10 mM stock solutions and stored in -20°C. Saponin, TMPD (N,N,N′,N′-tetramethyl-p-phenylenediamine), and sodium ascorbate were obtained from Sigma-Aldrich, (St. Louis, MO, USA).

Kovalenko I., Glasauer A., Schöckel L., Sauter D.R., Ehrmann A., Sohler F., Hägebarth A., Novak I, & Christian S. (2016). Identification of KCa3.1 Channel as a Novel Regulator of Oxidative Phosphorylation in a Subset of Pancreatic Carcinoma Cell Lines. PLoS ONE, 11(8), e0160658.

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Endothelin-1 and NS309 Protocols

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Endothelin-1 and NS309 were purchased from Sigma-Aldrich (St. Louis, MO).

Liu Y., Cole V., Lawandy I., Ehsan A., Sellke F.W, & Feng J. (2018). Decreased coronary arteriolar response to KCa channel opener after cardioplegic arrest in diabetic patients. Molecular and Cellular Biochemistry, 445(1), 187-194.

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Embryoid Body-Based Differentiation Using bSF Medium

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Cells were mechanically detached from the conventional 6-well plate culture (Nunc) using a cell scraper and transferred to 6-well suspension plates (Greiner) in a well-per-well scheme for EB-based differentiation in basic serum-free (bSF) medium (Kempf et al., 2011 (link), Xu et al., 2008 (link)) consisting of DMEM (Gibco) supplemented with 1% non-essential amino acids (Gibco), 2 mmol/L L-glutamine (Gibco), 0.1 mmol/L β-mercaptoethanol (Gibco), 5.6 mg/L transferrin (Sigma-Aldrich), and 37.2 μg/L sodium selenite (Sigma-Aldrich). bSF medium was supplemented with 5 μM SB203580 (Graichen et al., 2008 (link)), a p38 MAPK inhibitor, from day 0 to day 6. The compounds 1-ethyl-2-benzimidazolinone (EBIO; 1 M stock solution; Tocris Bioscience or Institute of Organic Chemistry at Hannover University), CyPPA (15 mM stock solution, Tocris), and NS309 (3 mM stock solution, Tocris) were added at respective concentrations and timing as described in Results. DMSO (Sigma-Aldrich), the solvent of EBIO, CyPPA, and NS309, was applied in respective amounts in controls. Apamin was diluted in sterile H₂O (Tocris, 0.1 mM stock solution). Medium was changed every 2–3 days. Before EB dissociation for endpoint analysis, cells were cultured for 24 hr in bSF medium.

Jara-Avaca M., Kempf H., Rückert M., Robles-Diaz D., Franke A., de la Roche J., Fischer M., Malan D., Sasse P., Solodenko W., Dräger G., Kirschning A., Martin U, & Zweigerdt R. (2017). EBIO Does Not Induce Cardiomyogenesis in Human Pluripotent Stem Cells but Modulates Cardiac Subtype Enrichment by Lineage-Selective Survival. Stem Cell Reports, 8(2), 305-317.

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Isolated Vessel Experiments with aCSF

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Isolated vessel experiments were performed using aCSF (mM): NaCl 122.0, NaHCO₃ 26.0, KCl 3.0, NaH₂PO₄ 1.25, MgCl₂ 1.0, CaCl₂ 2.0, and glucose 4.0. Buffer solutions were made each week and stored without glucose at 4 °C. Glucose was added prior to each experiment. Zero calcium aCSF was made without the addition of CaCl₂. U46619, NS309, L-NNA, and SNP were purchased from Sigma (St. Louis, MO, USA). Stock solutions were made weekly and stored at 4° C for L-NNA and SNP. Aliquots of U46619 and NS309 were made and kept frozen at −20° C until use.

Chan S.L., Nelson M.T, & Cipolla M.J. (2018). Transient Receptor Potential Vanilloid-4 Channels Are Involved in Diminished Myogenic Tone in Brain Parenchymal Arterioles in Response to Chronic Hypoperfusion in Mice. Acta physiologica (Oxford, England), 225(2), e13181.

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Evaluating Inflammatory Modulators

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HCQ, LPS, nigericin, ATP, NS169, NS309, clotrimazol, IBTX, TEA and BAPTA-AM were from Sigma (St Louis). Anti-IL-1beta and anti-caspase 1 were from Santa Cruz Biotechnologies. Human IL-1beta ELISAs were from BD.

Eugenia Schroeder M., Russo S., Costa C., Hori J., Tiscornia I., Bollati-Fogolín M., Zamboni D.S., Ferreira G., Cairoli E, & Hill M. (2017). Pro-inflammatory Ca++-activated K+ channels are inhibited by hydroxychloroquine. Scientific Reports, 7, 1892.

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Modeling Dopaminergic Neurons using MESC2.10 Cells

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These cells are a clone of MESC2.10 cells, studied and validated previously.^{14 (link), 15 (link)} After 4–6 days of in vitro differentiation, cells expressed the DAT, the VMAT-2, TH, and the neuronal form of β-III tubulin.^{15 (link), 45 (link)} The compounds NS309, rotenone and apamin^{21 (link)} were obtained from Sigma, Deisenhofen, Germany. NS309 is a selective positive modulator for SK1-3/IK channels, with an absolute requirement for a minimum concentration of intracellular Ca²⁺. It equally changes the sensitivity of SK1, SK2 or SK3 channels towards calcium and it is devoid of any effect on BK type channels.^{19 (link)} NS309 was added simultaneously with the rotenone treatment.

Dolga A.M., de Andrade A., Meissner L., Knaus H.G., Höllerhage M., Christophersen P., Zischka H., Plesnila N., Höglinger G.U, & Culmsee C. (2014). Subcellular expression and neuroprotective effects of SK channels in human dopaminergic neurons. Cell Death & Disease, 5(1), e999-.

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Vascular Smooth Muscle Contractility

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Adenosine, NS309, KCl, and papaverine were purchased from Sigma Aldrich (St. Louis, MO, USA). Apamin and TRAM-34 were purchased from Tocris (Minneapolis, MN, USA). Diltiazem was purchased from MP Biomedicals (Santa Ana, CA, USA). Adenosine stock solution was made every other day, and stock solutions of papaverine and Diltiazem were made weekly and stored at 4°C until use. NS309 and TRAM-34 were diluted in DMSO, and Apamin in double-distilled H₂O, and stock solutions aliquoted and stored at −20 °C until use. Isolated PA experiments were performed using aCSF containing (mM): NaCl 122.0, NaHCO₃ 26.0, NaH₃PO₄ 1.25, KCl 3.0, MgCl₂ 1.0, CaCl₂ 2.0, and glucose 4.0. aCSF with higher concentrations of KCl (8 – 40 mM) were made with reduced amounts of NaCl to maintain constant osmolality. Buffer solutions were made each week and stored without glucose at 4 °C. Glucose was added immediately prior to each experiment. aCSF was aerated with 5 % CO₂, 10 % O₂ and 85 % N₂ to maintain pH at 7.40 ± 0.05 and the temperature within the arteriograph chamber bath was maintained at 37.0 ± 0.2 °C throughout the experiments.

Johnson A.C, & Cipolla M.J. (2018). Impaired Function of Cerebral Parenchymal Arterioles in Experimental Preeclampsia. Microvascular research, 119, 64-72.

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Vasodilation Assay with Diltiazem and KCl

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Diltiazem was purchased from MP Biomedicals (Santa Ana, CA, United States), and all other compounds, including NS309, KCl, SNP, papaverine, and those used to make aCSF were purchased from Sigma Aldrich (St. Louis, MO, United States). Stock solutions of SNP, papaverine and Diltiazem were made weekly and stored at 4°C until use. NS309 stock solution was aliquoted and stored at -20°C until use. aCSF contained (mM): NaCl 122.0, NaHCO₃ 26.0, NaH₃PO₄ 1.25, KCl 3.0, MgCl₂ 1.0, and CaCl₂ 2.0. aCSF with higher concentrations of KCl (5, 10, 15, 20, 30, 40 mM) were made with reduced amounts of NaCl to maintain constant osmolality. Buffer solutions were made each week and stored without glucose at 4°C. Glucose was added (4.0 mM) immediately prior to each experiment. Zero calcium aCSF was made similarly, omitting the CaCl₂ and with the addition of 0.5 mM EGTA. aCSF was aerated with 5% CO₂, 10% O₂ and 85% N₂ to maintain pH at 7.40 ± 0.05 and the temperature within the arteriograph chamber was maintained at 37.0 ± 0.1°C throughout the experiments.

Johnson A.C., Tremble S.M, & Cipolla M.J. (2022). Experimental Preeclampsia Causes Long-Lasting Hippocampal Vascular Dysfunction and Memory Impairment. Frontiers in Physiology, 13, 889918.

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Pharmacological Modulation of Cellular Signaling

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ADP, apamin, endothelin-1, indomethacin, L-NAME, NS309, SNP,
substance P, and TRAM34 were purchased from Sigma-Aldrich (St Louis,
Mo).

Zhang Z., Shi G., Liu Y., Xing H., Kabakov A.Y., Zhao A.S., Agbortoko V., Kim J., Singh A.K., Koren G., Harrington E.O., Sellke F.W, & Feng J. (2020). Coronary endothelial dysfunction prevented by small-conductance calcium-activated potassium channel activator in mice and patients with diabetes. The Journal of thoracic and cardiovascular surgery, 160(6), e263-e280.

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