Mk 8776

Manufactured by Selleck Chemicals

Sourced in United States, China

The MK-8776 is a laboratory equipment product designed for scientific research and experimentation. It serves as a tool for conducting various analyses and measurements. The core function of the MK-8776 is to provide accurate and reliable data to support the research process, without making any interpretations or extrapolations about its intended use.

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Lab products found in correlation

36 protocols using mk 8776

Mitotic Spindle Inhibitor Efficacy

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Drugs, suppliers, and concentrations used were Barasertib (Aurora B inhibitor; alternative name AZD1152‐HQPS; SelleckChem S1147; 1.11 nM); CHR‐6494 (Haspin inhibitor; MedChem Express HY‐15217; 500 nM); CW069 (HSET inhibitor; SelleckChem S7336; 25.0 μM); Etoposide (Topoisomerase II inhibitor; SelleckChem S1225; 333 nM); GSK461364 (PLK1 inhibitor; SelleckChem S2193; 2.20 nM); GSK923295 (CENP‐E inhibitor; SelleckChem S7090; 3.20 nM); Ispinesib (KIF11 inhibitor; alternative name SB‐715992; SelleckChem S1452; 1.70 nM); MK‐5108 (Aurora A inhibitor; alternative name VX‐689; SelleckChem S2770; 0.576 nM); MK‐8776 (CHK1 inhibitor; alternative name SCH 900776; SelleckChem S2735; 9.00 nM); Paclitaxel (microtubule inhibitor; SelleckChem S1150; 2.67 nM); Vinblastine (microtubule inhibitor; Sigma V1377; 2.40 nM); and YM155 (BIRC5 inhibitor; SelleckChem S1130; 0.540 nM).

Maier L.J., Kallenberger S.M., Jechow K., Waschow M., Eils R, & Conrad C. (2018). Unraveling mitotic protein networks by 3D multiplexed epitope drug screening. Molecular Systems Biology, 14(8), e8238.

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Lung Cancer Cell Line Characterization

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NSCLC cell lines A549, HCC4006 and Calu-1 (three human lung adenocarcinoma cell lines), and SK-MES-1 and KLN205 (two human lung squamous carcinoma cell lines) were obtained from the Shanghai Institute for Biological Science (China). All cell lines were purchased between 2012 and 2015 and authenticated based on growth rate, morphology, and viability and were frequently confirmed to be mycoplasma free. The cells were cultured in RPMT 1640 media supplemented with 10% heat-inactivated fetal calf serum (FBS), L-glutamine and 100 U/ml penicillin, and 100 μg/ml streptomycin sulfate. The cells were maintained in a humidified incubator in 5% CO₂ at 37 °C.
Antibodies to various antigens were as follows: CHK1, PCHK1 (S296), PCHK1 (S317), PCHK1 (S345), CHK2, PCHK2 (T68), PCHK2 (S516), Cdc25A, PCdc25C (S216), H2AX, γH2AX (S139), PH3 (S10histone), ATM, PATM (S1981), DNA-PKCs, PDNA-PKCs (S2056), and GAPDH were from Cell Signaling. FANCL, FANCD2, BRCA2, PARP1, RAD51, caspase-3, cleaved caspanse-3, PARP and cleaved PARP from Santa Cruz. Gemcitabine was from Hanson Pharmaceutic (China), Cisplatin from Yangtze River Pharmaceutic (China), MK-8776 from Selleck Chemicals, CHK2 inhibitor II from Abcam. Gemcitabine was dissolved in PBS. Cisplatin, MK-8776 and CHK2 inhibitor II were dissolved in DMSO and used at the specified concentrations.

Dai C.H., Wang Y., Chen P., Jiang Q., Lan T., Li M.Y., Su J.Y., Wu Y, & Li J. (2017). Suppression of the FA pathway combined with CHK1 inhibitor hypersensitize lung cancer cells to gemcitabine. Scientific Reports, 7, 15031.

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Synthesis and Acquisition of Compounds

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CCT244747 was synthesised as described [29 (link)]. MK8776 was purchased from (S2735 Selleckchem, U.K.). Etoposide was purchased from (E1383, Scientific Lab Supplies Nottingham, U.K.), SN38 (2684) and Mitomycin C (3258) were obtained from (R&D systems, Abingdon, U.K.). All other compounds were purchased from Sigma–Aldrich.

Hunter J.E., Campbell A.E., Butterworth J.A., Sellier H., Hannaway N.L., Luli S., Floudas A., Kenneth N.S., Moore A.J., Brownridge P.J., Thomas H.D., Coxhead J., Taylor L., Leary P., Hasoon M.S., Knight A.M., Garrett M.D., Collins I., Eyers C.E, & Perkins N.D. (2022). Mutation of the RelA(p65) Thr505 phosphosite disrupts the DNA replication stress response leading to CHK1 inhibitor resistance. Biochemical Journal, 479(19), 2087-2113.

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Long-Term Drug Exposure Assay

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Short-term (72 h drug exposure) was performed as previously described^{21 (link)}.
For long-term exposure, cells were seeded at low density in a six-well plate. Treatment started 24 h later, and refreshed twice weekly until 80% confluency. Cells were stained using crystal violet (Sigma-Aldrich) after fixation using 2% paraformaldehyde (Sigma-Aldrich, Zwijndrecht, The Netherlands).
KU-60019, Wortmannin, ETP-46464, VE-821, MK-8776 (SCH 900776), PF-477736, LY2603618/Rabusertib, and Palbociclib were purchased from Selleckchem (Munich, Germany), LY2606368/Prexasertib from Medchem (Sollentuna, Sweden). Adavosertib (MK-1775) from Biovision (Milpitas, USA). All were dissolved in a DMSO stock dilution (10 mM), except Palbociclib (10 mM in ddH₂O). Assays contained <1% DMSO. All drug-survival assays depict the standard error of the mean (SEM) of three independent experiments in triplicate.

van Harten A.M., Buijze M., van der Mast R., Rooimans M.A., Martens-de Kemp S.R., Bachas C., Brink A., Stigter-van Walsum M., Wolthuis R.M, & Brakenhoff R.H. (2019). Targeting the cell cycle in head and neck cancer by Chk1 inhibition: a novel concept of bimodal cell death. Oncogenesis, 8(7), 38.

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Cell Line-Based Anticancer Drug Evaluation

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AZD6738 and VE-822 were purchased from MedKoo Biosciences (Morrisville, NC, USA), MK-8776 and LY2603618 from Selleckchem (Munich, Germany), and MMC and 5-FU from Sigma-Aldrich (Hamburg, Germany). Oxaliplatin and carboplatin were kindly donated from the cytostatic drug department of the University Hospital Marburg. AZD6738, used in the xenograft in vivo assay, was purchased from AdooQ Bioscience (Irvine, CA, USA).

Job A., Tatura M., Schäfer C., Lutz V., Schneider H., Lankat-Buttgereit B., Zielinski A., Borgmann K., Bauer C., Gress T.M., Buchholz M, & Gallmeier E. (2020). The POLD1R689W variant increases the sensitivity of colorectal cancer cells to ATR and CHK1 inhibitors. Scientific Reports, 10, 18924.

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Optimized Cell Death Pathway Analysis

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The following commercial reagents were used in this study: TNFα (Novoprotein, C008), 5Z-7-Oxozeaenol (5Z-7) (Sigma-Aldrich, O9890), TPCA-1 (MCE, HY-10074), PH-797804 (MCE, HY-10403), LY2228820 (MCE, HY-13241), MK2i (MCE, HY-12834A), GSK’872 (Calbiochem, 530389), NSA (Necrosulfonamide) (Calbiochem, 480073), MK-8776 (Selleck, SCH900776), PD-407824 (Sigma-Aldrich, PZ0111), BHT (Sigma-Aldrich, 34750), BHA (Sigma-Aldrich, 20021), AP20187 (MCE, HY-13992). R-7-Cl-O-Nec-1(Nec-1s), zVAD.fmk and SM164 were made by custom synthesis. Final concentrations of the compounds used in all experiments: TNFα, 20 ng/mL; zVAD, 25 μM; SM164, 50 nM; Nec-1s, 10 μM.

Zu R., Yu Z., Zhao J., Lu X., Liang W., Sun L., Si C., Zhu K., Zhang T., Li G., Zhang M., Zhang Y., Liu N., Yuan J, & Shan B. (2021). Quantitative analysis of phosphoproteome in necroptosis reveals a role of TRIM28 phosphorylation in promoting necroptosis-induced cytokine production. Cell Death & Disease, 12(11), 994.

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Inhibitors Targeting Cellular Signaling

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XL413 (S7547), BMS265246 (S2014), ON-01910 (S1362), PD0166285 (S8148), LDC000067 (S7461), PF-03814735 (S2725), D 4476 (S7642), VE-821 (S8007), AZD8055 (S1555), AZD2014 (S2783), AZD6738 (S7693) and MK-8776 (2735) were purchased from Selleck Chemicals. THZ531 (A8736) was purchased from ApexBio. XL413 (205768), BLU9931 (206192) and LY3177833 (206762) were purchased from MedKoo. TAK-931 (CT-TAK931) was purchased from Chemietek. XL413 (A13677) was also purchased from AdooQ BIOSCIENCE. The SHP2 inhibitor used in this study is covered by a patent application (WO 2015/107495A1; compound #57) and was synthesized as described previously^{23 (link)}.
Antibodies against HSP90 (sc-7947, sc-13119), p53 (sc-126), p21 (sc-6246), and SHP2 (sc-280) were purchased from Santa Cruz Biotechnology. Antibodies against CDC7 (ab77668), p-MCM2 (ab109133, ab133243), MCM2 (ab4461), p-SHP2 (ab62322), PCNA (ab2426), and Cleaved caspase-3 (ab2303) were purchased from Abcam. Antibodies against γH2AX (#9718), p-S6RP (#4856, #5364), S6RP (#2317), p-4EBP1 (#9456, #2855, #9455), 4EBP1 (#9644), p-IGF-1R/INSR (#3024), IGF-1R (#9750), p-PDGFRβ (#3161), PDGFRβ (#4564), p-AKT (#4060), and AKT (#2920) were purchased from Cell Signalling. EGFR antibody (610017) was purchased from BD Biosciences. H3K9Me3 antibody (49-1008) and p-EGFR (44-788) were from Thermo Fisher Scientific.

Wang C., Vegna S., Jin H., Benedict B., Lieftink C., Ramirez C., Leite de Oliveira R., Morris B., Gadiot J., Wang W., du Chatinier A., Wang L., Gao D., Evers B., Jin G., Xue Z., Schepers A., Jochems F., Mulero Sanchez A., Mainardi S., te Riele H., Beijersbergen R.L., Qin W., Akkari L, & Bernards R. (2019). Inducing and exploiting vulnerabilities for the treatment of liver cancer. Nature, 574(7777), 268-272.

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Squamous Cell Carcinoma Cell Lines

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Primary squamous carcinoma cell lines (University of Turku-Squamous cell carcinoma (UT-SCC)) have been established previously in the Grenman lab, Turku University, Finland and FaDu, Cal33, SAS, HSC4 and XF354 were from ATCC/LGC (Wesel, Germany). Cells were cultivated in DMEM supplemented with 10% FCS, 2 mM glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C at 10% CO₂. For the determination of cellular survival cells were treated with up to 1.5 μg/mL to mitomycin C (Medac, Wedel, Germany) or 1 µM topotecan (Sigma, St. Louis, MS, USA,) for 6 h or 24 h and irradiation experiments were performed using radiation doses up to 6 Gy at a dose rate of 1.2 Gy/min using a Gulmay X-ray machine (GULMAY Medical, Byfleet, UK) or UV 5mJ (BioRad Laboratories, Berkeley, CA, USA). All treatments were performed in 37 °C and 10% CO₂ atmosphere. CHK1 inhibition was achieved by using siGENOME human CHEK1 siRNA (Horizon Discovery, Cambridge, UK) up to 150 nM for 24 h or the small molecule inhibitor MK8776 (Selleck Chemicals, Houston, TX, USA) at 2 µM for 2 h.

Bold I.T., Specht A.K., Droste C.F., Zielinski A., Meyer F., Clauditz T.S., Münscher A., Werner S., Rothkamm K., Petersen C, & Borgmann K. (2021). DNA Damage Response during Replication Correlates with CIN70 Score and Determines Survival in HNSCC Patients. Cancers, 13(6), 1194.

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Nitazoxanide and Chk1 Inhibitors Protocol

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Nitazoxanide (NTZ) and the cell cycle checkpoint kinase 1 (Chk1) inhibitors MK-8776 and Prexasertib HCl were purchased from Selleck Chemicals (Selleck, Houston, TX, USA). The anti-VP2 monoclonal antibody was purchased from Ingenasa (Ingenasa, Madrid, Spain). Beta-actin monoclonal antibody was purchased from Thermo Fisher (Thermo Fisher, Waltham, MA, USA). The ProteinExt^® Mammalian Total Protein Extraction Kit was purchased from TransGen (TransGen Biotech, Beijing, China). Total RNA extraction was conducted using the RNeasy Mini Kit (QIAGEN, Hilden, Germany). The reverse transcription kit FastKing RT Kit (With gDNase) and quantitative PCR kit SuperReal PreMix Plus (SYBR Green) were purchased from TIANGEN (TIANGEN, Beijing, China).

Su X., Zhou H., Han Z., Xu F., Xiao B., Zhang J., Qi Q., Lin L., Zhang H., Li S, & Yang B. (2024). Transcriptional Differential Analysis of Nitazoxanide-Mediated Anticanine Parvovirus Effect in F81 Cells. Viruses, 16(2), 282.

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Chemotherapy Agents and Targeted Inhibitors

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Chemotherapy drugs (Doxorubicin, Ara-C, and Daunorubicin) were obtained from the clinical pharmacy at Cincinnati Children’s Hospital. WEE1 inhibitor MK-1775 and CHK1 inhibitor MK-8776 were obtained from Selleckchem.
Detailed methods are described in the Supplementary information.

Dong Y., Zhao X., Feng X., Zhou Y., Yan X., Zhang Y., Bu J., Zhan D., Hayashi Y., Zhang Y., Xu Z., Huang R., Wang J., Zhao T., Xiao Z., Ju Z., Andreassen P.R., Wang Q.F., Chen W, & Huang G. (2019). SETD2 mutations confer chemoresistance in acute myeloid leukemia partly through altered cell cycle checkpoints. Leukemia, 33(11), 2585-2598.

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