Carboplatin

Manufactured by Teva Pharmaceuticals

Sourced in Spain

Carboplatin is a platinum-based antineoplastic agent used in the treatment of various types of cancer. It functions as a cytotoxic agent, inhibiting DNA synthesis and cell division. Carboplatin is commonly used in combination with other chemotherapeutic drugs as part of cancer treatment regimens.

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Lab products found in correlation

Paclitaxel, by Merck Group (4 mentions) Irinotecan, by Pfizer (2 mentions) Cisplatin, by Pfizer (2 mentions) Cantharidin, by Bio-Techne (2 mentions) Etoposide, by Fresenius (2 mentions) Lb 100, by Selleck Chemicals (2 mentions) Biological reagents, by Merck Group (2 mentions) Infinite m200 pro, by Tecan (2 mentions) Skov3, by American Type Culture Collection (2 mentions) Infinite m200 pro plate reader, by Tecan (1 mentions) Celltiter glo 3d cell viability assay, by Promega (1 mentions) Doxycycline, by Merck Group (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Cytochrome c, by Cell Signaling Technology (1 mentions) Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions) Tom20, by Santa Cruz Biotechnology (1 mentions) Rad50, by Cell Signaling Technology (1 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Rad51, by Santa Cruz Biotechnology (1 mentions)

18 protocols using carboplatin

Carboplatin Dose-Response Assay

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Cells were seeded in a 96-well plate (2,000 cells per well) and incubated overnight before treatment with sequential dilutions of carboplatin (Teva Pharmaceuticals; range, 50–1,000 μmol/L) for 6 (ovarian cancer cell lines) or 7 (patient-derived ovarian cancer cells) days. Cell viability was assessed with MTS reagent that contains a tetrazolium compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron-coupling reagent (phenazine methosulfate; PMS). The optical density (absorbance at 490 nm) was measured with an Infinite M200 Pro plate reader (Tecan, Inc.). IC₅₀ values were determined in GraphPad Prism (GraphPad Software, Inc.).

Compadre A.J., van Biljon L.N., Valentine M.C., Llop-Guevara A., Graham E., Fashemi B., Herencia-Ropero A., Kotnik E.N., Cooper I., Harrington S.P., Kuroki L.M., McCourt C.K., Hagemann A.R., Thaker P.H., Mutch D.G., Powell M.A., Sun L., Mosammaparast N., Serra V., Zhao P., Lomonosova E., Khabele D, & Mullen M.M. (2023). RAD51 Foci as a Biomarker Predictive of Platinum Chemotherapy Response in Ovarian Cancer. Clinical Cancer Research, 29(13), 2466-2479.

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Organoid Carboplatin Sensitivity Assay

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For organoid carboplatin sensitivity assay, 20,000 cells per 10 μL of 75% Cultrex were plated into the wells of a black walled 96-well plate. On day 2, media containing 1, 5, 10, 25, 50, and 75 μmol/L of carboplatin (Teva; No. 00703424601) were added to the organoids. On day 7, an equal volume of CellTiter-Glo 3D Cell Viability Assay (Promega; No. G9681) was added to each well, and the luminescence was read using a Tecan plate reader. The percentage of cell viability was calculated and graphed using Microsoft Excel and GraphPad Prism.

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Anticancer Drug Formulation Protocol

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Gemcitabine hydrochloride was obtained from SANDOZ (Novartis Division, Italia), Trabectedin from PharmaMar (Pharma Mar, S.A., Madrid, Spain), 5-FU and Carboplatin from TEVA (Teva Italia srl, Milano, Italy), and Oxaliplatin from SUN Ranbaxy (Sun Pharmaceutical Industries Ltd., Goregaon, Mumbai, India). Paclitaxel and Doxycycline were obtained from Sigma Aldrich (Sigma–Aldrich, St. Louis, MO, USA). All drugs were dissolved in water for injection and aliquoted in different working solutions to avoid degradation.

Varamo C., Peraldo-Neia C., Ostano P., Basiricò M., Raggi C., Bernabei P., Venesio T., Berrino E., Aglietta M., Leone F, & Cavalloni G. (2019). Establishment and Characterization of a New Intrahepatic Cholangiocarcinoma Cell Line Resistant to Gemcitabine. Cancers, 11(4), 519.

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Tarextumab and Chemotherapy in Allograft Mice

Cited 1 time

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NOD/SCID mice implanted with OMP-LU66 or TKO allografts were randomized and treated with a control antibody or tarextumab (OMP-59R5, 40mg/kg, Q2W) as a single agent or in combination with the chemotherapy agents carboplatin (25 mg/kg, QW, Teva) and irinotecan (25 mg/kg, QW, Pfizer). We used carboplatin and irinotecan (instead of cisplatin and etoposide) for these longer term studies as they are less toxic, better tolerated by the mice, and have been shown to have similar efficacies as cisplatin and etoposide^{36 (link),37 (link)}. To avoid the side effects of total Notch pathway inhibition in vivo^{38 (link),39 (link)}, we sought to reduce Notch signaling with the Notch2/3 antagonist tarextumab. After approximately four cycles, chemotherapy was discontinued and tarextumab dosing was continued until study completion. Mice with tumor volumes at or exceeding the 2500mm^{3 (link)} limit permitted by the IACUC were sacrificed regardless of timepoint.

Lim J.S., Ibaseta A., Fischer M.M., Cancilla B., O’Young G., Cristea S., Luca V.C., Yang D., Jahchan N.S., Hamard C., Antoine M., Wislez M., Kong C., Cain J., Liu Y.W., Kapoun A.M., Garcia K.C., Hoey T., Murriel C.L, & Sage J. (2017). Intratumoral heterogeneity generated by Notch signaling promotes small cell lung cancer. Nature, 545(7654), 360-364.

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DNA Retardation Assay with Compounds

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Different concentrations of the test compounds (2.5, 10.0, 20.0, 50.0 and 100.0 μM) were incubated with 0.1 μg of plasmid DNApHOT1 (TopoGen, Buena Vista, CO, USA) at 37 °C for 30 min in 10.0 mM Tris-HCl (pH 7.9). Carboplatin (TEVA, Tel Aviv, Israel) was used as a positive control of the retardation of DNA migration. The reaction products were separated in 1% agarose gel electrophoresis under a maximal voltage of 1.8 V/cm for 5 h. The gel was then stained with 0.5 μg/mL ethidium bromide. The DNA contained in the gel was visualized via UV fluorescence with a ChemiDoc MP Imaging system (Bio-Rad Laboratories, Hercules, CA, USA) at a wavelength of 254 nm.

Moiseeva N., Eroshenko D., Laletina L., Rybalkina E., Susova O., Karamysheva A., Tolmacheva I., Nazarov M, & Grishko V. (2023). The Molecular Mechanisms of Oleanane Aldehyde-β-enone Cytotoxicity against Doxorubicin-Resistant Cancer Cells. Biology, 12(3), 415.

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Iron Compounds Preparation for Cell Proliferation

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Iron compounds were first reconstituted in DMSO and then diluted with sterile and bidistilled water to obtain 100 µM stock solutions with a concentration of 4% DMSO (Sigma-Aldrich, Burlington, MA, USA). Just before the experiments, these solutions were further diluted with sterile complete medium to obtain the desired final concentrations. In the cell proliferation experiments, cisplatin (Pfizer, Madrid, Spain) and carboplatin (Teva, Madrid, Spain) were included as controls.

Castro J., Bravo M., Albertí M., Marsal A., Alonso-De Gennaro M.J., Martínez-Ferraté O., Claver C., van Leeuwen P.W., Romero I., Benito A, & Vilanova M. (2022). Dinuclear Iron Complexes of Iminopyridine-Based Ligands as Selective Cytotoxins for Tumor Cells and Inhibitors of Cancer Cell Migration. Pharmaceutics, 14(12), 2801.

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TNBC Cell Line Maintenance and Treatment

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Three TNBC cell lines (Hs578t, MDA-MB-231, and SUM159) were purchased from the American Type Culture Collection (Manassas, VA, USA). All cells were maintained in Dulbecco’s Modified Eagle’s Medium (Invitrogen, Grand Island, NY) with 10% fetal bovine serum (Thermos Scientific Hyclone, Rockford, IL) in a humidified 5% CO₂ incubator at 37°C. Chloroquine diphosphate salt (Sigma-Aldrich, St. Louis, MO) and carboplatin (Teva Pharmaceuticals, Sellersville, PA) were dissolved in cell culture-grade water at the concentration of 10 mM. CD marker antibodies (BD Biosciences, San Jose, California), MitoSOX Red Mitochondrial Superoxide Indicator (Thermo Fisher Scientific, Waltham, MA), Cell Proliferation Reagent WST-1 (Clontech, Mountain View, CA) were acquired. Cytochrome C, LC-3B, p62, NQO1, Rad50, PARP and β-actin antibodies were purchased from Cell Signaling Technology (Danvers, MA). Rad51, β-tubulin, and Tom20 antibodies were purchased from Santa Cruz Biotechnology Inc. Dallas, TX. γ-H₂AX (EMD Millipore, Billerica, MA) and bcl-2 antibodies (BioLegend, San Diego, CA), were purchased from their respective companies.

Liang D.H., Choi D.S., Ensor J.E., Kaipparettu B.A., Bass B.L, & Chang J.C. (2016). The autophagy inhibitor chloroquine targets cancer stem cells in triple negative breast cancer by inducing mitochondrial damage and impairing DNA break repair. Cancer letters, 376(2), 249-258.

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Optimal Dosing for Intraperitoneal Chemotherapy Regimen

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In this study, we chose an intraperitoneal route for administration of chemotherapy because it is less demanding to administrate in mice. Moreover, in human studies intraperitoneal delivery has at least the same response rate as the intravenous route and therefore are used more and more in clinical practice [26] (link). To determine the maximum tolerable dosage (MTD) of carboplatin (Teva, Helsingborg, Sweden 10 mg/ml) and paclitaxel (Fresenius Kabi, Halden, Norway 6 mg/ml), the following different dosages were evaluated. carboplatin 15, 20 or 30 mg/kg, paclitaxel 12, 16 or 20 mg/kg as monotherapy or combined (n = 3 mice per group, total 27 mice) twice weekly for three consecutive weeks (Q2Wx3). Body weight was monitored for 28 days. A combination consisting of carboplatin 15 mg/kg together with paclitaxel 12 mg/kg was found to be the MTD. At the end of study the mice were euthanized.

Helland Ø., Popa M., Vintermyr O.K., Molven A., Gjertsen B.T., Bjørge L, & McCormack E. (2014). First In-Mouse Development and Application of a Surgically Relevant Xenograft Model of Ovarian Carcinoma. PLoS ONE, 9(3), e89527.

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Biological Reagents and Treatments

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Biological reagents were purchased from Sigma-Aldrich (St. Louise, MO) or Thermo Fisher Scientific (Waltham, MA). Electrophoresis and western blotting reagents were purchased from Bio-Rad (Irvine, CA). LB100 was purchased from SelleckChem (Houston, TX), cantharidin from Tocris (Minneapolis, MN), carboplatin was purchased from Teva Pharmaceuticals (Irvine, CA) and etoposide was purchased from Fresenius Kabi (Schaumburg, IL).

Mirzapoiazova T., Xiao G., Mambetsariev B., Nasser M.W., Miaou E., Singhal S.S., Srivastava S., Mambetsariev I., Nelson M.S., Nam A., Behal A., Atri P., Muschen M., Tissot F.L., Miser J., Kovach J.S., Sattler M., Batra S.K., Kulkarni P, & Salgia R. (2021). Protein Phosphatase 2A as a Therapeutic Target in Small Cell Lung Cancer. Molecular Cancer Therapeutics, 20(10), 1820-1835.

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Synergistic Anti-Cancer Drug Evaluation

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Cells (5,000 per well) were plated in each well of a 96-well plate and incubated overnight. Cells were then serum starved and treated with 0.7–1000 nM paclitaxel (Sigma-Aldrich), 0.01–108 μM carboplatin (Teva Pharmaceuticals, Sellersville, PA), 1–5 μM AVB-500 (Aravive), or a combination of these drugs. carboplatin and AVB-500 were dissolved in saline. paclitaxel was dissolved in DMSO for all experiments. After 72 hours, MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) solution (Promega, Madison, WI) was added to the cells (20μL/well), incubated for 2 hours, and then the absorbance was measured at 490 nm with a 96-well plate reader (Tecan infinite M200 Pro). The drug combination index (CI) (20 (link)) was calculated using CompuSyn software to assess for synergy between drugs..

Mullen M.M., Lomonosova E., Toboni M.D., Oplt A., Cybulla E., Blachut B., Zhao P., Noia H., Wilke D., Rankin E.B., Kuroki L.M., Hagemann A.R., Hagemann I.S., McCourt C.K., Thaker P.H., Mutch D.G., Powell M.A., Mosammaparast N., Vindigni A, & Fuh K.C. (2021). GAS6/AXL inhibition enhances ovarian cancer sensitivity to chemotherapy and PARP inhibition through increased DNA damage and enhanced replication stress. Molecular cancer research : MCR, 20(2), 265-279.

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