Otx 015

Manufactured by MedChemExpress

Sourced in United States, China

OTX-015 is a small-molecule inhibitor that targets the bromodomain and extra-terminal (BET) family of proteins, including BRD2, BRD3, BRD4, and BRDT. It is a tool compound used in research to study the role of BET proteins in various biological and disease processes.

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6 protocols using otx 015

Solvent Preparation for In Vitro and In Vivo Studies

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OTX-015 and LY2606368 (MedChem Express, Monmouth Junction, NJ, USA) and SRA737 (Chemietek, Indianapolis, IN, USA) were dissolved in 100% dimethyl sulfoxide (DMSO) for in vitro studies. The final concentration of DMSO for all drugs in all cell cultures was ≤0.1%. OTX-015 and SRA737 were dissolved in 10% DMSO, 20% PEG400, 5% Tween 80, and 65% water for the in-vivo studies.

Pandya P.H., Cheng L., Saadatzadeh M.R., Bijangi-Vishehsaraei K., Tang S., Sinn A.L., Trowbridge M.A., Coy K.L., Bailey B.J., Young C.N., Ding J., Dobrota E.A., Dyer S., Elmi A., Thompson Q., Barghi F., Shultz J., Albright E.A., Shannon H.E., Murray M.E., Marshall M.S., Ferguson M.J., Bertrand T.E., Wurtz L.D., Batra S., Li L., Renbarger J.L, & Pollok K.E. (2020). Systems Biology Approach Identifies Prognostic Signatures of Poor Overall Survival and Guides the Prioritization of Novel BET-CHK1 Combination Therapy for Osteosarcoma. Cancers, 12(9), 2426.

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Evaluation of BRD4 Inhibitors in Cells

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Dimethyl sulfoxide (DMSO) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The BRD4 inhibitors, JQ1, I-BET151 and OTX015, were all purchased from MedChemExpress (Shanghai, China), dissolved in DMSO, and stored in small aliquots – 20 ℃.

Pang Y., Bai G., Zhao J., Wei X., Li R., Li J., Hu S., Peng L., Liu P, & Mao H. (2022). The BRD4 inhibitor JQ1 suppresses tumor growth by reducing c-Myc expression in endometrial cancer. Journal of Translational Medicine, 20, 336.

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HNSCC Cell Line Inhibitor Screening

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The present study utilized human HNSCC cell lines SCC25 and CAL27, along with human embryonic kidney-derived 293 T cells, which were procured from the American Type Culture Collection (ATCC). The SCC25 cells were cultivated in DMEM/F12, supplemented with 10% fetal bovine serum (FBS, #086–1500, WISENT) and 400 ng/mL hydrocortisone. On the other hand, CAL27 and 293 T cells were maintained in DMEM supplemented with 10% FBS, and all cells were incubated under constant conditions at 37 °C with 5% CO₂. The small molecule inhibitors THZ1 (#V2557) and JQ1 (#V0411) were obtained from InvivoChem, whereas OTX-015 (#202,590–98-5) and CPI-637 (#1,884,712–47-3) were purchased from MedChemExpress. Upon reaching an approximate fusion rate of 60–70% overnight, the culture medium was replaced with varying concentrations of small molecule inhibitors for continuous cell culture and subsequent investigations.

Cai H., Liang J., Jiang Y., Wang Z., Li H., Wang W., Wang C, & Hou J. (2024). KLF7 regulates super-enhancer-driven IGF2BP2 overexpression to promote the progression of head and neck squamous cell carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 43, 69.

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Cytotoxicity Assay of JQ-1, OTX-015, and ZBC-260

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Cells were seeded in 96-well plates at a density of 5000 cells per well and then incubated with dimethyl sulfoxide (DMSO; control) or a series of 2-fold-diluted concentrations of JQ-1 (cat. no. HY-13030, MedChemExpress), OTX-015 (cat. no. HY-15743, MedChemExpress), and ZBC-260 for 4 days. Three replicates were made for each concentration of the drugs. After incubation, Cell Titer-Glo luminescent assays (cat. no. G7572, Promega) were performed to measure cell viability following the manufacturer’s protocol.

Shi C., Ye Z., Han J., Ye X., Lu W., Ji C., Li Z., Ma Z., Zhang Q., Zhang Y., He W., Chen Z., Cao X., Shou X., Zhou X., Wang Y., Zhang Z., Li Y., Ye H., He M., Chen H., Cheng H., Sun J., Cai J., Huang C., Ye F., Luo C., Zhou B., Ding H, & Zhao Y. (2020). BRD4 as a therapeutic target for nonfunctioning and growth hormone pituitary adenoma. Neuro-Oncology, 22(8), 1114-1125.

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Multiplexed Cytotoxicity Assay Protocol

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ML324 (HY-12725), A196 (HY-100201), Irinotecan (HY-16562), Doxorubicin (HY-15142), EPZ-6438 (HY-13803), GSK503 (HY-12856), 5-Azacytidine (HY-10586), JQ-1 (HY-13030), OTX-015 (HY-15743), I-BET151 (HY-13235) and wortmannin (HY-10197) were from MedChemExpress. Lipofectamine 3000 (L3000015), CellMask Green (C37608) and CellMask Deep Red (C10046) Plasma Membrane Stain were from Invitrogen.

Wang J., Li G.L., Ming S.L., Wang C.F., Shi L.J., Su B.Q., Wu H.T., Zeng L., Han Y.Q., Liu Z.H., Jiang D.W., Du Y.K., Li X.D., Zhang G.P., Yang G.Y, & Chu B.B. (2020). BRD4 inhibition exerts anti-viral activity through DNA damage-dependent innate immune responses. PLoS Pathogens, 16(3), e1008429.

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Characterization of MYC-driven Medulloblastoma Cell Lines

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MB cell lines D-283 (MYC-amplified) and D-341(MYC-amplified) were purchased from American Type Culture Collection. HD-MB03 (MYC-amplified) MB cell line was purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (Germany). ONS-76 (non-MYC-amplified, SHH) MB cell line was purchased from Sekisui-XenoTech (USA). Cell lines were authenticated by their respective resources using short tandem repeat profiling. All cell lines were also tested for negative mycoplasma contamination using MycoSensor-PCR assay kit (Agilent-Technologies). All cell lines were cultured and maintained using RPMI-1640 media supplemented with 10% heat-inactivated FBS, penicillin (100 U/ml) and streptomycin (100 µg/ml) (Life Technologies) in a humidified incubator at 5% CO2 and 95% air atmosphere at 37 °C. Experiments were performed using less than 10 passages for each cell line. The BET-protein inhibitors JQ1 and OTX-015 and pan-HDAC inhibitor panobinostat were purchased from MedChemExpress LLC.

Kling M.J., Kesherwani V., Mishra N.K., Alexander G., McIntyre E.M., Ray S., Challagundla K.B., Joshi S.S., Coulter D.W, & Chaturvedi N.K. (2022). A novel dual epigenetic approach targeting BET proteins and HDACs in Group 3 (MYC-driven) Medulloblastoma. Journal of Experimental & Clinical Cancer Research : CR, 41, 321.

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