Mek162

Manufactured by Merck Group

Sourced in United States

MEK162 is a laboratory equipment product manufactured by Merck Group. It is designed for specific research applications, but a detailed description of its core function cannot be provided in a concise, unbiased, and factual manner without the risk of interpretation or extrapolation.

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Lab products found in correlation

3 protocols using mek162

Evaluating Combination Inhibitor Efficacy

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H1975, H460, and A549 cells were treated with 0–3 μM MEK162 (MEK inhibitor, Sigma, St. Louis, MO, USA) or 0–15 μM BKM120 (PI3K inhibitor, Sigma), alone or in combination for 48 h. H1975, H460, and A549 cells were treated with 1 μM BIBW2992 (EGFR inhibitor, Sigma), a second-generation EGFR-TKI, or 2 μM ARQ197 (MET inhibitor, Sigma) combination for 48 h. A stock solution of the chemical inhibitors was prepared in 100% DMSO and further diluted with normal saline. The inhibitors were used at a final concentration of 0.05% DMSO.

Qu G.P., Shi M., Wang D., Wu J.H., Wang P., Gong M.L, & Zhang Z.J. (2021). Dual targeting of MEK and PI3K effectively controls the proliferation of human EGFR-TKI resistant non-small cell lung carcinoma cell lines with different genetic backgrounds. BMC Pulmonary Medicine, 21, 208.

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Quantification of Neutrophil Extracellular Traps

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NETs were quantified essentially as described with some modifications (32 (link)). Neutrophils were attached to poly D-lysine coated 96-well black transparent bottom plates (Thermo Scientific, Rochester, NY) in assay medium containing 10 μM Sytox Orange (Life Technologies, Grand Island, NY, USA). For inhibitor tests, cells were pretreated with specific inhibitors for 15 minutes prior to bacterial infection. Neutrophils were infected with 1 to 50 MOI Pseudomonas aeruginosa PA14 for dose-dependent extracellular DNA release assay or 10 MOI for inhibitor experiments. Fluorescence (excitation: 530 nm, emission: 590 nm) was measured in a fluorescence plate reader (Varioskan Ascent, Thermo Scientific) for up to 5 hours at 37 °C. Fluorescence of samples containing 500 μg/mL saponin (Sigma, St. Louis, MO) with neutrophils was used as the maximal signal for DNA release (100%). Rise in fluorescence was referred to as “extracellular DNA release” and expressed as relative fluorescence units (RFU) or normalized on saponin-treated as maximal signal (referred to as “% of max”). The following inhibitor concentrations were used: DPI (diphenylene iodonium, NADPH oxidase inhibitor, 10 μM, Sigma), U0126 (MEK1/2 inhibitor, 25 μM, Sigma), MEK162 (MEK1/2 inhibitor, 50 μM, Sigma), PD98059 (MEK1 inhibitor, 20 μM, Sigma) and cytochalasin-D (cytoskeleton inhibitor, 10 μM, Sigma).

Yoo D.G., Winn M., Pang L., Moskowitz S.M., Malech H.L., Leto T.L, & Rada B. (2014). RELEASE OF CYSTIC FIBROSIS AIRWAY INFLAMMATORY MARKERS FROM PSEUDOMONAS AERUGINOSA-STIMULATED HUMAN NEUTROPHILS INVOLVES NADPH OXIDASE-DEPENDENT EXTRACELLULAR DNA TRAP FORMATION. Journal of immunology (Baltimore, Md. : 1950), 192(10), 4728-4738.

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Establishing Neuroblastoma and Other Cell Lines

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SHSY-5Y human neuroblastoma cells were purchased from ATCC (CRL-2266) and cultured as a monolayer at 37 °C, 5% CO₂ in Dulbecco’s Modified Eagles Medium (DMEM)/F12 containing 10% fetal bovine serum (FBS), and penicillin/streptomycin. HEK293, NIH3T3, AtT20 and PC-12 were cultured in DMEM with 10% FBS, all reagents from Life Technologies, Inc (Grand Island, NY, USA). All cell cultures were detached with trypsin and transferred to new 75 cm² culture flasks (Fisher, Pittsburgh, PA, USA) once weekly. Human recombinant bFGF, EGF, Metformin, MEK-162, Imatinib and Sunitinib were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA). The growth factors and Metformin were dissolved in media while the other compounds were dissolved in DMSO. Small aliquots were stored at −80 °C and used after a single freeze–thaw cycle.

Zhang D., Yong W.H., Movassaghi M., Rodriguez F.J., Yang I., McKeever P., Qian J., Li J.Y., Mao Q., Newell K.L., Green R.M., Welsh C.T, & Heaney A.P. (2022). Whole Exome Sequencing Identifies PHF14 Mutations in Neurocytoma and Predicts Responsivity to the PDGFR Inhibitor Sunitinib. Biomedicines, 10(11), 2842.

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