Nms p715

Manufactured by Merck Group

Sourced in United States

The NMS-P715 is a laboratory equipment product manufactured by Merck Group. It is a precision instrument designed for scientific research and analysis applications. The core function of the NMS-P715 is to provide accurate and reliable measurements, but no further details on its intended use can be provided in an unbiased and factual manner.

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Lab products found in correlation

7 protocols using nms p715

Microscopy Imaging Methodology with Inhibitors

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Inhibitors were dissolved in DMSO and were used at the following concentrations: 50 nM CENP-E inhibitor (GSK923295, Cambridge Bioscience), 2–3 μM MPS1 inhibitor (NMS-P715, Merck Chemicals) and 0.15–0.45 μM reversine (Sigma-Aldrich).
Antibodies were used as follows: mouse α-tubulin (Thermo Fisher Scientific, 32-2500), 1:5000 (western blotting or WB) and 1:500 (immunofluorescence or IF); rabbit α-tubulin (Abcam, ab52866), 1:500 (IF); mouse CENP-A (Abcam, ab13939), 1:200 (IF); human CREST (Antibodies Incorporated, 15-234-0001), 1:300 (IF); rabbit pericentrin (Abcam, ab4448), 1:2000 (IF); rabbit KIF11 (Novus Biologicals, NB5000-181), 1:1000 (IF); mouse p53 (Cell Signaling Technologies, 48818S), 1:1000 (IF); and rabbit MAD2 (Millipore UK, MABE866), 1:1000 (WB) and 1:500 (IF).

Dale K.L., Armond J.W., Hynds R.E, & Vladimirou E. (2022). Modest increase of KIF11 expression exposes fragilities in the mitotic spindle, causing chromosomal instability. Journal of Cell Science, 135(17), jcs260031.

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Chemical Inhibitors for Cellular Assays

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The chemical inhibitors used in this study were centrinone (150 nM; LCR-263; synthesized by Sudia MediTech; Wong et al., 2015 (link)); MDM2 inhibitor ((2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5-dimethyl-4,5-dihydro-1H-imidazol-1-yl)(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)methanone); 1 µM; synthesized by Sudia MediTech; Ding et al., 2009 ); doxorubicin (1 µM; Sigma-Aldrich); cytochalasin D (4 µM; Sigma-Aldrich); monastrol (100 µM; Tocris Bioscience); nocodazole (2.5 µg/ml; Sigma-Aldrich); and NMS-P715 (2 µM; EMD Millipore).

Meitinger F., Anzola J.V., Kaulich M., Richardson A., Stender J.D., Benner C., Glass C.K., Dowdy S.F., Desai A., Shiau A.K, & Oegema K. (2016). 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration. The Journal of Cell Biology, 214(2), 155-166.

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Inhibition of Mitotic Regulators in HeLa Cells

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HeLa cells were cultured in Dulbecco's modified Eagle's Media (DMEM) supplemented with 10% fetal calf serum (FCS) and antibiotics, penicillin and streptomycin). For inhibition studies, cells were treated with 10 µM monastrol (1305, Tocris) for Eg5 inhibition for 3 h or 200 nM NMS-P715 (475949-5MG, Merck) for MPS1 inhibition prior to filming or fixation. 2ME2 (Sigma) was added directly to DMEM containing FCS (Invitrogen). Cells were transfected with siRNAs as described [30 (link)]. All siRNA oligos used against TAO1 are listed in the electronic supplementary material. For Control-St siRNA treatment, negative control (12935-300, Invitrogen) was used. All siRNA transfections were performed using Oligofectamine (Life Technologies) according to the manufacturer's instructions. For tetracycline induction, cells were treated with 1 ng ml^–1 tetracycline (Sigma).

Shrestha R.L., Tamura N., Fries A., Levin N., Clark J, & Draviam V.M. (2014). TAO1 kinase maintains chromosomal stability by facilitating proper congression of chromosomes. Open Biology, 4(6), 130108.

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Chemical Compound Dissolution Protocol

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Cpd-5, MPI-0479605 (ApexBio, Houston, TX, USA), NMS-P715 (Merck Millipore), reversine (Sigma), Mps1-IN-3 (Sigma) and AZ3146 (Axon Medchem, Groningen, Netherlands) were all dissolved in DMSO.

Koch A., Maia A., Janssen A, & Medema R.H. (2015). Molecular basis underlying resistance to Mps1/TTK inhibitors. Oncogene, 35(19), 2518-2528.

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Pharmacological Inhibitors in Cell Research

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Reversine was obtained from Cayman Chemical or Sigma-Aldrich and used at a working concentration of 0.5 μM or 2μM; Monastrol (working concentration 100 μM) from Tocris; SB203580 (working concentration 10 μM) from CellSignalingTechnology; Thymidine (working concentration 5 mM), Aphidicolin (working concentration 400 nM), RO-3306 (working concentration 7.5 μM), Chk2 inhibitor II hydrate (working concentration 10 μM), VE821 (working concentration 1 μM) and Nocodazole (working concentration 330 nM) were obtained from Sigma-Aldrich. NMS-P715 (working concentration 1 μM) was obtained from EMD/Millipore.

Santaguida S., Richardson A., Iyer D.R., M’Saad O., Zasadil L., Knouse K.A., Wong Y.L., Rhind N., Desai A, & Amon A. (2017). Chromosome mis-segregation generates cell cycle-arrested cells with complex karyotypes that are eliminated by the immune system. Developmental cell, 41(6), 638-651.e5.

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Chemical Agents for Cell Biology Research

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Doxycycline was used at 1 μg/ml (Sigma), Indole-3-acetic acid (IAA) used at 500 μM (Sigma, I5148), CDK1 inhibitor RO-3306 used at 9 μM (Sigma-Aldrich, SML0569), NMS-P715 0.8 μM (Millipore Sigma, 475949), ICRF-193 used at 100 nM (Enzo life sciences, BML-GR332–0001) S-Trityl-L-cysteine (STLC) used at 5 μM (Enzo Life Sciences, ALX-105–011-M500), dTAG-13 used at 500 nM (Sigma-Aldrich, SML2601), Okadaic acid used at 500 nM (Fisher Scientific, 11–362-5U), Geneticin^® Selective Antibiotic (G418 Sulfate) used at 300 μg/ml (Gibco, 10131035).

Broberger C., Johansen J., Johansson C., Schalling M, & Hökfelt T. (1998). The neuropeptide Y/agouti gene-related protein (AGRP) brain circuitry in normal, anorectic, and monosodium glutamate-treated mice. Proceedings of the National Academy of Sciences of the United States of America, 95(25), 15043-15048.

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Inhibition of TBK1, TTK, and APC/C in Cell Studies

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Cells were treated with BX795 (#s1274, selleckchem), MRT67307 (#SML0702, Millipore Sigma) or amlexanox (#4857, Tocris) to inhibit TBK1 and NMS-P715 (#475949, Millipore Sigma) to inhibit TTK Plasmids expressing GST-Cdh1 and GST-Cdc20 were kind gift from Dr. Lixin Wan. Phospho (S172) TBK1 (#5483), phospho PLK1 (#5472S), phospho histone H3 (#3377), TTK (#3255T), APC1 (#13329), pAurora A (Thr288), B (Thr232), C (Thr 198) (#2914S) and total TBK1 (#3013S) antibodies were purchased from Cell Signaling, and antibody against Cdc20 (SC-13162) was purchased from Santa Cruz Biotechnology. β-actin (#A1978, clone AC-15) and α-tubulin (#T6074) antibodies were purchased from Sigma Chemical Co. Cyclin B1 antibody (#5472S) was purchased from BD Biosciences. pTTK antibody (#44–1325G) was purchased from Novex. BubR1 (ab70544), Mad2 (ab97777) and Cdh1 (ab89535) antibodies were purchased from Abcam. γ-tubulin antibody (# PA5–34815) was purchased from Invitrogen. Control siRNA (sc-37007) was purchased from Santa Cruz Biotechnology and TBK1 siRNA (# 4457298, ID: s761) was purchased from Ambion.

Maan M., Agrawal N.J., Padmanabhan J., Leitzinger C.C., Rivera-Rivera Y., Saavedra H.I, & Chellappan S.P. (2020). Tank Binding Kinase 1 modulates spindle assembly checkpoint components to regulate mitosis in breast and lung cancer cells. Biochimica et biophysica acta. Molecular cell research, 1868(3), 118929.

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