Bay1895344

Manufactured by Selleck Chemicals

BAY1895344 is a laboratory chemical product. It is a solid substance that is typically used in research and development applications. The core function of this product is to serve as a chemical compound for analysis and experimentation purposes in a laboratory setting.

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6 protocols using bay1895344

Preparation of Inhibitor Stock Solutions

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All compounds were reconstituted to 10 mM stock using DMSO (Sigma Aldrich) and stored at −20 °C. AXL inhibitor R428 (BGB324) (#S2841), ATR inhibitor BAY1895344 (#S8666), ATR inhibitor VE-821 (#S8007), PARP1 inhibitor Olaparib (#S1060), PARP1 inhibitor Niraparib (#S2741) were purchased from Selleckchem.

Yeo X.H., Sundararajan V., Wu Z., Phua Z.J., Ho Y.Y., Peh K.L., Chiu Y.C., Tan T.Z., Kappei D., Ho Y.S., Tan D.S., Tam W.L, & Huang R.Y. (2023). The effect of inhibition of receptor tyrosine kinase AXL on DNA damage response in ovarian cancer. Communications Biology, 6, 660.

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Anticancer Drug Screening in Organoids

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Organoids were seeded into 96-well plates at 1,000 cells/well (Day 0). The indicated concentrations of Rucaparib (Selleckchem, S1098), Niraparib (MCE, HY-10619), Olaparib (Selleckchem, S1060), Gemcitabine (MCE, HY-B0003), Doxorubin (sigma, D1515), Paclitaxel (Selleckchem, S1150), Carboplatin (Sigma, 1096407), Seliciclib (MCE, HY-30237), PHA767491(Sigma, PZ0178), BAY1895344 (Selleckchem, S8666), Chloroquine (Selleckchem, S4157) and YKL-5–124 (a gift from Dr. Kwok-kin Wong) were added on the day following seeding (Day 1). Media were changed, and fresh drug was added on Day 3. Cell viability was assessed on Day 5 by adding10 μl PrestoBlue and incubating for 30 min in 37℃. Fluorescence was measured in a FlexStation® 3 Multi-Mode Microplate Reader (BOSTONind). Results were normalized to DMSO controls, and IC₅₀ values were determined using Graphpad Prism 7.

Zhang S., Iyer S., Ran H., Dolgalev I., Gu S., Wei W., Foster C.J., Loomis C.A., Olvera N., Dao F., Levine D.A., Weinberg R.A, & Neel B.G. (2020). Genetically Defined, Syngeneic Organoid Platform for Developing Combination Therapies for Ovarian Cancer. Cancer discovery, 11(2), 362-383.

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Cell Viability Assay of MEL290 Cells

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To determine cell viability, MEL290 cells were seeded in 96-well plates at a density of 1000 cells per well and treated with 2.5 nM BAY-1895344 (Selleck, S8666) or 2 μM VE-822. After incubation with 10 μL CCK-8 reagent (Dojindo Laboratories, Kumamoto, Japan) per well, the absorbance was measured at a wavelength of 450 nm at the indicated time points. The data were recorded and analyzed. The quantifications presented were obtained from three independent biological replicates.

Zong C., Zhang Z., Gao L., He J., Wang Y., Li Q., Liu X., Yang J., Chen D., Huang R., Zheng G., Jin X., Wei W., Jia R, & Shen J. (2023). APOBEC3B coordinates R-loop to promote replication stress and sensitize cancer cells to ATR/Chk1 inhibitors. Cell Death & Disease, 14(6), 348.

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BRCA1/2 Complex Characterization and Inhibition

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Primary antibodies used are as follows: Myc (9E10, Covance), BRCA1 (D9, sc-6954, Santa Cruz), BRCA1-pS1423 (sc-101647, Santa Cruz), BRCA2 (OP95, EMD Millipore), BARD1 (H300, Santa Cruz), Abraxas (ab139191, AbCam), RAD51 (H-92, Santa Cruz), GAPDH (FL-335, Santa Cruz) and β-Actin (AC-15, Santa Cruz). Anti-human PALB2 was described before (22 (link)). Anti-mouse BRCA1 was a gift from Dr. Andre Nussenzweig (NCI). Key chemicals used are Olaparib (S1060, Selleckchem), cisplatin (S1166, Selleckchem), ATM inhibitors KU-55933 (S1092, Selleckchem), KU-60019 (S1570, Selleckchem), AZD1390 (S8680, Selleckchem); ATR inhibitors VE-821 (S8007, Selleckchem), AZ20 (S7050, Selleckchem), VE-822 (S7102, Selleckchem), and BAY-1895344 (S8666, Selleckchem).

Foo T.K., Vincelli G., Huselid E., Her J., Zheng H., Simhadri S., Wang M., Huo Y., Li T., Yu X., Li H., Zhao W., Bunting S.F, & Xia B. (2021). ATR/ATM-mediated phosphorylation of BRCA1 T1394 promotes homologous recombinational repair and G2/M checkpoint maintenance. Cancer research, 81(18), 4676-4684.

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Evaluating Synergistic Effects of Targeted Therapies

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For the case study, MCF-7 and SOX11 overexpression SH-EP cell line were grown in RPMI medium supplemented with 10% FCS and 2mM L-Glutamine and 100 IU/mL penicillin/streptomycin. The generation of SOX11 overexpression is described in [24 (link)]. To evaluate synergism and the dose-response relationship, cells were seeded in a 384-well plate (Corning COS3764), at a density of 3x10³ cells per well. Cells were allowed to adhere overnight, after which these were exposed to the respective treatment. The drugs used were: AZD2281 (Selleckchem, S1060), MK-1775 (Selleckchem, S1525), prexasertib (MedChem Express, HY-18174A) and BAY 1895344 (Selleckchem, S8666). The treatment was performed by the D300 TECAN. Cell proliferation was monitored for 72h, in which pictures were taken through IncuCyte S3 Live Cell Imaging System each 2h. Each image was analysed through the IncuCyte S3 Software. Cell proliferation was monitored by analysing the occupied area (% confluence) over time.

Nunes C., Anckaert J., De Vloed F., De Wyn J., Durinck K., Vandesompele J., Speleman F, & Vermeirssen V. (2024). HTSplotter: An end-to-end data processing, analysis and visualisation tool for chemical and genetic in vitro perturbation screening. PLOS ONE, 19(1), e0296322.

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ATM/ATR Inhibitors in CRISPR Editing

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Where indicated, the culture medium was supplied with 1 μM ATM inhibitor (KU55933), 0.5 μM ATR inhibitor (AZ20), 1 μM ATR inhibitor (VE821) or 0.5 μM ATR inhibitor (BAY1895344) (Selleckchem) one hour prior to gRNA transfection.

Korsholm L.M., Gál Z., Lin L., Quevedo O., Ahmad D.A., Dulina E., Luo Y., Bartek J, & Larsen D.H. (2019). Double-strand breaks in ribosomal RNA genes activate a distinct signaling and chromatin response to facilitate nucleolar restructuring and repair. Nucleic Acids Research, 47(15), 8019-8035.

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