Bml 277

Manufactured by Selleck Chemicals

Sourced in United States

The BML-277 is a laboratory instrument designed for the analysis and characterization of chemical compounds. It is a versatile tool that can be used for a variety of applications in research and development laboratories. The core function of the BML-277 is to provide accurate and reliable data on the physical and chemical properties of samples, enabling researchers to make informed decisions about their work.

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Lab products found in correlation

7 protocols using bml 277

Evaluating Drug Sensitivity in Leukemia Cells

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For drug-sensitivity assays, primary BM pro-B/pre-B cells, WT Em-Myc lymphoma cells or Hoxa9-Meis1 AML cells, or their derivatives, were seeded into 96-well plates at a density of 1 Â 10 5 cells/well and treated with serial diluted drugs (5-point 1:4 dilution), including olaparib, niraparib (Selleckchem, S2741), veliparib (Sellechchem, S1004), S63845, etoposide (Selleckchem, S1225), RG-7388. Cell death was assessed at 48 hours using propidium iodide (PI; Sigma, P4864) uptake, or PI/AnnexinV-APC (WEHI mAb lab), measured on an LSRIIW flow cytometer (BD Biosciences). For CellTiter-Glo assays, 1 Â 10 4 cells were seeded into white 96-well plate (Greiner, 655083) and cell viability was assessed using the CellTiter-Glo Luminescent Assay (Promega, G9241).
To test the interaction between olaparib and BML-277 (Selleckchem, S8632), primary pro-B/pre-B cells (1 Â 10 4 ), Em-Myc lymphoma cells (1 Â 10 4 ), or human cancer cell lines (2 Â 10 3 ) were seeded into 96-well white plates and treated with both drugs, either individually, or in a combination matrix (each drug 5-point 1:4 dilution, olaparib horizontal dilution, BML-277 vertical dilution). Cell viability was determined at 48 hours (pro-B/pre-B or Em-Myc) or at 96 hours (human cancer cell lines) using Cell Titer Glo Luminescent Assay. Bliss scores were calculated as described previously (36) .

Xu Z., Vandenberg C.J., Lieschke E., Di Rago L., Scott C.L, & Majewski I.J. (2021). CHK2 Inhibition Provides a Strategy to Suppress Hematologic Toxicity from PARP Inhibitors. Molecular cancer research : MCR, 19(8).

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Compound Preparation and Storage

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Fisetin, dactolisib, GSK1059615, BML-277, and WNK463 were purchased from Selleckchem. Quercetin was purchased from Sigma-Aldrich. Stock solutions of the compounds were prepared on the day of an experiment by dissolving the powder in dimethyl sulfoxide (DMSO). Stock solutions were kept at − 80 °C for long-term storage. Working concentrations were diluted from respective stock solutions using complete growth medium. Dilutions of Fisetin and Quercetin were prepared in phenol red-free media. All solutions were protected from light.

Shahi Thakuri P., Gupta M., Singh S., Joshi R., Glasgow E., Lekan A., Agarwal S., Luker G.D, & Tavana H. (2020). Phytochemicals inhibit migration of triple negative breast cancer cells by targeting kinase signaling. BMC Cancer, 20, 4.

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Comprehensive Cancer Cell Panel Experiments

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The cell lines used in experiments included MFE-296, HEC50B, DU4475, BT-474, HCC1954, MDA-MB-468, SK-OV-3, KM12, HCC2998, SW480, HCT15. They were obtained through the MD Anderson Characterized Core Cell Line Facility and the MDACC CRC Moon Shot program. Cell lines were authenticated by fingerprinting using short tandem repeat testing. The absence of mycoplasma contamination was also verified. Cells were maintained in RPMI-1640 supplemented with 10% heat-inactivated FBS and 1% penicillin with streptomycin. The targeted compounds (PARP inhibitor – Olaparib, Chk2 inhibitor – BML-277, AKT inhibitor MK-2206, MET inhibitor – JNJ-38877605, HER2 inhibitor – Neratinib, AURKA inhibitor – Alisertib, CDK4/6 inhibitor – Ribociclib, MAP3K4 inhibitor – Doramapimod (directly inhibiting downstream signaling partner, p38/MAPK14), PIK3CA inhibitor – Buparlisib) were obtained from Selleckchem.

Li X., Dowling E.K., Yan G., Dereli Z., Bozorgui B., Imanirad P., Elnaggar J.H., Luna A., Menter D.G., Pilié P.G., Yap T.A., Kopetz S., Sander C, & Korkut A. (2022). Precision combination therapies based on recurrent oncogenic co-alterations. Cancer discovery, 12(6), 1542-1559.

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Glioblastoma Cell Lines and Primary Culture

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Glioblastoma cell lines U373, LN229, and T98G were available from the American Tissue Type Culture Collection (USA). KUGBM8 primary cell line was established by Dr. Filiz Şenbabaoğlu from patient samples in collaboration with Koç University Hospital Neurosurgery Department; ethical approval for KUGBM8 cell line was obtained from the Koç University Institutional Review Board (2014.079.IRB2.022) [48 (link)]. Protocol of primary cell line generation was adapted from [69 (link)]. All parental and irradiated cell populations cells were cultured in DMEM (Gibco, Gaithersburg, MD, USA) supplemented with 10% FBS (Gibco, Gaithersburg, MD, USA) and 1% Penicillin-Streptomycin (Gibco, Gaithersburg, MD, USA). All cells were maintained at 37 °C in a humidified incubator with 5% CO₂. To achieve hypoxic conditions, cells were maintained at 37 °C in a humidified incubator with 5% CO₂ and 1% O₂. AZD7762 (Selleckchem, S1532), AZD6738 (Ceralasertib, Selleckchem, S7693), LY2603618 (Rabusertib, Selleckchem, S2626), KU55933 (Selleckchem, S1092), BML-277 (Selleckchem, S8632), Bleomycin (Selleckchem, S1214), and Temozolomide (Selleckchem, S1237) were used for drug treatment experiments.

Pinarbasi-Degirmenci N., Sur-Erdem I., Akcay V., Bolukbasi Y., Selek U., Solaroglu I, & Bagci-Onder T. (2022). Chronically Radiation-Exposed Survivor Glioblastoma Cells Display Poor Response to Chk1 Inhibition under Hypoxia. International Journal of Molecular Sciences, 23(13), 7051.

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Culturing Hepatocellular Carcinoma Cell Lines

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The human HCC cell lines (HepG2 and Hep3B) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in MEM (Gibco, Carlsbad, MD, USA) and DMEM (Gibco), respectively, and were supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin. They were maintained at 37 °C in a humidified atmosphere with 5% CO₂. Cabozantinib was obtained from Sigma-Aldrich (St. Louis, MO, USA) and BML-277 was purchased from Selleck Chemicals (Houston, TX, USA).

Jeon Y., Kim T., Kwon H., Kim J.K., Park Y.T., Ham J, & Kim Y.J. (2023). Cannabidiol Enhances Cabozantinib-Induced Apoptotic Cell Death via Phosphorylation of p53 Regulated by ER Stress in Hepatocellular Carcinoma. Cancers, 15(15), 3987.

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Carboplatin and BML-277 Dissolution Protocol

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Carboplatin (AdipoGen, San Diego, CA, USA, AG-CR1-3591) and BML-277 (SelleckChem, Houston, TX, USA, Cat#S8632) were dissolved in water (10 mg/mL) and in dimethyl sulfoxide (10 mM), respectively, and stored at 20 °C. Further dilutions of each drug for the respective experiments were freshly prepared in the growth medium. Table 1 lists antisera and dilutions used for various experiments.

Park D., Gharghabi M., Reczek C.R., Plow R., Yungvirt C., Aldaz C.M, & Huebner K. (2022). Wwox Binding to the Murine Brca1-BRCT Domain Regulates Timing of Brip1 and CtIP Phospho-Protein Interactions with This Domain at DNA Double-Strand Breaks, and Repair Pathway Choice. International Journal of Molecular Sciences, 23(7), 3729.

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Chemogenomic Screen of BML-277 in Cells

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BML-277 was purchased from Selleck (S8632), dissolved in dimethyl sulfoxide (DMSO), and stored at −80 °C in small aliquots. siRNAs were obtained from Sigma, and were transfected by jet-PRIME reagent (Polyplus) following the manufacturer’s instructions. The sequence of CHK2-siRNA is 5′-CCUGUGGAGAGGUAAAGCU-3′ and of control-siRNA is 5′-UUCUCCGAACGUGUCACGU-3′.

Tian Y., Wang Y., Xu S., Guan C., Zhang Q, & Li W. (2020). The Expression and Therapeutic Potential of Checkpoint Kinase 2 in Laryngeal Squamous Cell Carcinoma. Drug Design, Development and Therapy, 14, 2613-2622.

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