Nps2143

Manufactured by Merck Group

Sourced in Germany, United States

NPS2143 is a laboratory instrument designed for the measurement and analysis of various physical and chemical properties. It is a versatile piece of equipment that can be used in a wide range of scientific applications. The core function of NPS2143 is to provide accurate and reliable data for researchers and scientists.

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Lab products found in correlation

8 protocols using nps2143

Calcium Signaling in Leukemia Cells

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In order to test the effect of extracellular calcium on THP1 and K562 cells, calcium chloride was added to the cells in increasing concentrations (0.5, 1, 2.5, 5, 7.5 mM) prior to the adhesion and migration experiments. The cells were then incubated for 4 h at 37 °C.
To test the effect of the CaSR-antagonist NPS-2143 on THP1 cells, 1 × 10⁶ cells were treated with either vehicle (DMSO) or 5, 10 or 20 µM NPS-2143 (Sigma, Darmstadt, Germany) for 4 h at 37 °C.

Pereira R.S., Kumar R., Cais A., Paulini L., Kahler A., Bravo J., Minciacchi V.R., Krack T., Kowarz E., Zanetti C., Godavarthy P.S., Hoeller F., Llavona P., Stark T., Tascher G., Nowak D., Meduri E., Huntly B.J., Münch C., Pampaloni F., Marschalek R, & Krause D.S. (2023). Distinct and targetable role of calcium-sensing receptor in leukaemia. Nature Communications, 14, 6242.

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Calcium Signaling in Neonatal Rat Cells

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New born Wistar rats (3-day-old) were obtained from Academy of Military Medical Sciences (Tianjin, China). DMEM and fetal bovine serum (FBS) were from Gibco (USA) and HyClone (USA), respectively. Fura-2/AM was purchased from Biotium (USA). The rest of reagents, including trypsin, collagenase II, EGTA, DMSO, thapsigargin (TG), BAPTA-AM, TMB-8, 2-APB, BTP-2 (YM-58483), U73122, U73343, NPS2143, spermine, nifedipine and verapamil were purchased from Sigma-Aldrich (USA). CellTiter 96 AQueous One Solution Cell Proliferation Assay kit and CellTiter-Glo Luminescent Cell Viability Assay kit were purchased from Promega (USA).

Hu F., Pan L., Zhang K., Xing F., Wang X., Lee I., Zhang X, & Xu J. (2014). Elevation of Extracellular Ca2+ Induces Store-Operated Calcium Entry via Calcium-Sensing Receptors: A Pathway Contributes to the Proliferation of Osteoblasts. PLoS ONE, 9(9), e107217.

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Zebrafish Pharmacological Assay Protocol

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All drugs were made in 1000 X stock solution and stored in light-protected Eppendorf tubes at −20 °C. CNO (20 mmol/L; Tocris), NPS 2143 (20 mmol/L; Sigma), U0126 (10 mmol/L; LC laboratories), and rapamycin (2.5 mmol/L; Cayman) were dissolved in DMSO at the indicated concentrations. Zebrafish were exposed to 1 μM or 20 μM CNO, 1 μM NPS 2143, 10 μM U0126, and 2.5 µM rapamycin for 2 days, respectively, starting at 3 dpf and injected with EdU at 4 dpf. All drug stocks were dissolved in egg water within a petri dish (10 cm) to the working concentration. Larvae were fixed and stained at 5 dpf for α cell number and proliferation assays.

Gong Y., Yang B., Zhang D., Zhang Y., Tang Z., Yang L., Coate K.C., Yin L., Covington B.A., Patel R.S., Siv W.A., Sellick K., Shou M., Chang W., Danielle Dean E., Powers A.C, & Chen W. (2023). Hyperaminoacidemia induces pancreatic α cell proliferation via synergism between the mTORC1 and CaSR-Gq signaling pathways. Nature Communications, 14, 235.

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Investigating Calcium-Signaling Mechanisms

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Calcium acetate, CaSR antagonist NPS2143, calcium indicator Fluo 3-AM, and ERK kinase inhibitor U0126 were purchased from Sigma-Aldrich. Cell Counting Kit-8 (CCK-8) was purchased from Vazyme Biotech Co., Ltd. (Nanjing, China). DMEM/F12 and fetal bovine serum (FBS) were purchased from Gibco BRL. Polyclonal antibodies against β-actin, cyclin D1, ERK1/2, phospho-ERK1/2, and p21 were purchased from Cell Signaling Technology Inc. Polyclonal antibodies against CaSR were purchased from Abcam plc.

Ye J., Ai W., Zhang F., Zhu X., Shu G., Wang L., Gao P., Xi Q., Zhang Y., Jiang Q, & Wang S. (2016). Enhanced Proliferation of Porcine Bone Marrow Mesenchymal Stem Cells Induced by Extracellular Calcium is Associated with the Activation of the Calcium-Sensing Receptor and ERK Signaling Pathway. Stem Cells International, 2016, 6570671.

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Cinacalcet and NPS 2143 modulate LUAD cells

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Human LUAD A549 cell line was purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). CaSR agonist cinacalcet (catalog number: SML2012-50MG) and antagonist NPS 2143 (catalog number: SML0362-25MG) were purchased from Sigma-Aldrich. We treated A549 cells with cinacalcet (0, 0.3, 1, 3, and 10 μM) for 90 min, or NPS 2143 (0, 0.025, 0.05, 0.1, and 1 μM) for 24 h.

Li J., Liao P., Wang K., Miao Z., Xiao R., Zhu L, & Hu Q. (2020). Calcium Sensing Receptor Inhibits Growth of Human Lung Adenocarcinoma Possibly via the GSK3β/Cyclin D1 Pathway. Frontiers in Cell and Developmental Biology, 8, 446.

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In Vitro Inflammation Study Protocol

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For the in vitro inflammation studies, HAoECs were seeded into 24-well TPP tissue culture plates (MidSci, MO, USA) with a seeding density of 5 × 10⁴ cells/well. The cells were grown for 3—5 days until they reached 90% confluency. Confluent cells were washed with HEPES-buffered saline solution (PromoCell, C-40000) and changed to Q-media or low-serum medium (1% FBS) for 1 h. Cells were then pretreated with γ-EV for 2 h at different concentrations (0.01, 0.1, and 1 mM), followed by treatment with TNF-α (5 ng/mL) for another 6 h. Cell medium supernatants were collected for enzyme-linked immunosorbent assays (ELISAs) and for western blots, and cells were harvested with 2× hot Laemmli buffer followed by the addition of dithiothreitol (DTT) and boiled for 3—4 min. To confirm the role of the CaSR, NPS-2143 (1 μM) (Sigma, #SML0362) was used to block the CaSR activation for 1 h before the γ-EV pretreatment.

Guha S., Paul C., Alvarez S., Mine Y, & Majumder K. (2020). Dietary γ-Glutamyl Valine Ameliorates TNF-α-Induced Vascular Inflammation via Endothelial Calcium-Sensing Receptors. Journal of agricultural and food chemistry, 68(34), 9139-9149.

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Cell Viability Assay using Synthetic Peptides

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STC-1 cells were seeded into black 96-well plate (5 × 10 4 cells per well) and cell viability was measured as described Santos-Hernández et al. (2018) (link). The cells were exposed to synthetic peptides or GPCR antagonists for 2 h or 1.5h, respectively. After incubation, the medium was aspirated and it was incubated for 1 h with Alamar Blue solution 1:10; v:v, (AlamarBlue™ Cell Viability Reagent, ThermoFisher Scientific, Walthman, USA) prior fluorescence measurement at 590/530 excitation/emission wavelength. Synthetic peptides were tested in HEPES buffer (20 mM HEPES 1 M, 10 mM glucose, 140 mM NaCl, 4.5 mM KCl, 1.2 mM CaCl2, 1.2 mM MgCl2, pH 7.4), all reagents were supplied by Merck KGaA, Darmstadt, Germany. The CaSR antagonist, NPS2143 (Sigma-Aldrich, Sigma Aldrich-Merck KGaA, Darmstadt, Germany), and GPR93 antagonist, TC LPA5 4 (Tocris, Bio-Techne, Minneapolis, USA), were dissolved in dimethylsulfoxide (DMSO, Sigma Aldrich-Merck KGaA, Darmstadt, Germany) at 2% and then diluted with HEPES buffer (1:1, v:v). NPS2143 was tested at 100 µM, 50 µM, 25 µM, 12 µM and 6 µM; and TC LPA5 4 was tested at 8 µM, 4 µM and 2 µM. Cell viability experiments were performed in triplicate and the fluorescence measurement in duplicate.

Santos-Hernández M., Vivanco-Maroto S.M., Miralles B, & Recio I. (2023). Food peptides as inducers of CCK and GLP-1 secretion and GPCRs involved in enteroendocrine cell signalling. Food chemistry, 402.

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Investigate Gprc6a Signaling in RAW264.7 Cells

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RAW264.7 cells were seeded into 24-well plates at a density of with 1.5 × 10⁵ cells/ml and cultured for 12 h. The specific siRNA of Gprc6a (siRNA-2553 GCAGAAGACTAACACCAAA) was transfected into RAW264.7 cells for 24 h before reporter transfection and ucOCN treatment. The reporter plasmid pGL4.33[luc2P/SRE/Hygro] or pGL4.29 [luc2P/CRE/ Hygro] coupled with phRLTK was transfected into RAW264.7 cells followed by treatment with 100 ng/ml ucOCN, 20 mM L-ornithine (Sigma, USA) alone or combined with 30 µM NPS2143 (Sigma, USA) as previous reports desrption (Rueda et al., 2016 (link)) 36 h later. The measurement of luciferase activity was performed with the Dual Luciferase Reporter Assay kit (Promega, USA) following the manufactory’s instruction.

Wang H., Li J., Xu Z., Wu F., Zhang H., Yang C., Chen J., Ding B., Sui X., Guo Z., Li Y, & Dai Z. (2021). Undercarboxylated osteocalcin inhibits the early differentiation of osteoclast mediated by Gprc6a. PeerJ, 9, e10898.

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