Azd2461

Manufactured by Merck Group

Sourced in United States

AZD2461 is a laboratory equipment product manufactured by Merck Group. It is designed to perform specific scientific functions within a controlled laboratory environment. The core function of AZD2461 is to facilitate research and analysis, but further details about its intended use are not included in this factual and unbiased description.

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Lab products found in correlation

8 protocols using azd2461

Radiation Sensitivity of Colon Cancer Cells

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HCT116 (human colon cancer cell line, wtp53) and HCT116 p53-/-(human colon cancer cell line, p53 K/O) were a kind gift from B. Vogelstein, (Johns Hopkins University, Baltimore, MD).
While HT-29 (human colon cancer cell line, carrying R273H mutp53) were kindly provided by N Merendino (Tuscia University, Viterbo Italy). Cells were maintained in DMEM 1640 (Thermo Fisher Scientific) supplemented with 10% Fetal Bovine Serum (FBS) (Corning), L-glutamine, streptomycin (100 μg/ml) (Corning), and penicillin (100 U/ml) (Corning) (complete medium) in 5% CO 2 at 37 • C. Cells were always detached using Trypsin-EDTA solution (Biological Industries, Cromwell, CT, USA). Cells were plated in 6-well plates at a density of 2 × 10 5 cells/well in 2 ml and, the day after, treated with AZD2461 (30 μM) (Sigma Aldrich) for 24 h. In some experiment cells were plated in 6well plates as above reported and the day after pre-treated with AZD2461 (30 μM) (Sigma Aldrich) for 5 h, then irradiated with 1 Grey (1Gy) radiation and cultured for the next 24 h. Untreated cells were used as control. Irradiation was carried out using an ONCOR Impression Linear Accelerator (Siemens Medical Solutions USA, Inc, Concord, CA) at a dose rate of 1 Gy (95 UM/min).

Romeo M.A., Gilardini Montani M.S., Benedetti R., Arena A., Maretto M., Bassetti E., Caiazzo R., D'Orazi G, & Cirone M. (2021). Anticancer effect of AZD2461 PARP inhibitor against colon cancer cells carrying wt or dysfunctional p53. Experimental cell research, 408(2).

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Cell Culture Reagents and Inhibitors

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Dulbecco’s modified Eagle’s medium (DMEM, Cat# 10313-021), Z150 L-glutamine 100× (Cat# 25030-081), and Trypsin-EDTA (0.05%) phenol red (Cat# 25300062) were from Gibco by Life Technologies, Australia. SERANA fetal bovine serum (FBS) (Cat# FBS-AU-015, batch no. 18030416) was from Fisher Biotechnology, Australia. cOmplete, mini EDTA-free protease inhibitor cocktail (Cat# 11836170001), PhosSTOP Phosphatase Inhibitors (Cat# 4906837001), hydrogen peroxide 30% (w/w) solution (Cat# H1009), AZD2461 (Cat# SML 1858), and MG132 (Z-Leu-Leu-Leu-al, Cat# C2211) were from Sigma-Aldrich.

McMahon K.A., Stroud D.A., Gambin Y., Tillu V., Bastiani M., Sierecki E., Polinkovsky M.E., Hall T.E., Gomez G.A., Wu Y., Parat M.O., Martel N., Lo H.P., Khanna K.K., Alexandrov K., Daly R., Yap A., Ryan M.T, & Parton R.G. (2021). Cavin3 released from caveolae interacts with BRCA1 to regulate the cellular stress response. eLife, 10, e61407.

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Breast Cancer Cell Culture Protocol

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MCF-7, human breast (adenocarcinoma) cell line was sourced from Pasteur Institute of Iran and were grown in RPMI 1640 medium obtained from INOCLON (G. Innovative Biotech Co (INOCLON), Iran) supplemented with 10% FBS (Gibco, Rockville, MD, USA), antibiotic-antimycotic solution (containing 100 mg/ml of penicillin, 2.5 mg/L of amphotericin B, and 100 U/ml of streptomycin. 0.25% Trypsin-EDTA solution (G. Innovative Biotech Co (INOCLON), Iran) was used to detach cells from the surface. AZD2461, VPA, Tween 20, Trypan blue and Triton X-100 was procured from Sigma-Aldrich (St. Louis, MO, USA). Cell culture flasks and microtiter plates were supplied by Biofill (Jet Biofill, China). All other chemicals were of certified reagent analytical grade.

Sargazi S., Kooshkaki O., Zavar Reza J., Saravani R., Zarei Jaliani H., Mirinejad S, & Meshkini F. (2019). Mild antagonistic effect of Valproic acid in combination with AZD2461 in MCF-7 breast cancer cells. Medical Journal of the Islamic Republic of Iran, 33, 29.

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Evaluating Cell Proliferation by MTT Assay

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Cell proliferation was evaluated by MTT assay (Sigma Aldrich, Burlington, MA, USA). A total of 5 × 103 cells/well were plated in 96-well plates in 100 μL of complete medium. The day after, the cells were pre-treated with VPA (10 mM) and TSA (500 nM) for 1 h and then treated with AZD2461 (10 μM) (Sigma Aldrich, Burlington, MA, USA) for 72 h. Untreated cells were used as control. MTT assay was performed following manufacturer’s instruction. The plates were analyzed by Byonoy Absorbance 96 (Germany). The experiments were performed in triplicate and repeated three times.

Romeo M.A., Gilardini Montani M.S., Benedetti R., Arena A., D’Orazi G, & Cirone M. (2022). VPA and TSA Interrupt the Interplay between mutp53 and HSP70, Leading to CHK1 and RAD51 Down-Regulation and Sensitizing Pancreatic Cancer Cells to AZD2461 PARP Inhibitor. International Journal of Molecular Sciences, 23(4), 2268.

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Anti-Acanthamoeba Activity of PARP Inhibitors

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Obtained from TargetMol (Wellesley Hills, MA, USA), talazoparib, AZD2461, azilsartan, trovafloxacin, olaparib, and venadaparib were dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA). AZ9482 was obtained from MCE (MedChem Express, Monmouth Junction, NJ). Although talazoparib, AZ9482, AZD2461, azilsartan, and trovafloxacin were prepared at 10 mM concentrations, olaparib, venadaparib and AZ9482 were prepared at 200 mM concentrations. A. castellanii trophozoites were seeded at the log growth phase (10 4 trophozoites per well) in a 96-well white microplate (Eppendorf, Hamburg, Germany). Incubation with medium containing varied olaparib, venadaparib and AZ9482 concentrations (100, 200, 300, 400 , and 500 mM) was performed for 24 h, 48 h and 72 h at 26°C. Trophozoites viability assay results, derived from at least three wells per condition and compared with diluted DMSO, were

Chen L., Han W., Jing W., Feng M., Zhou Q, & Cheng X. (2024). Novel anti-Acanthamoeba effects elicited by a repurposed poly (ADP-ribose) polymerase inhibitor AZ9482. Frontiers in cellular and infection microbiology, 14.

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Evaluating Cellular Sensitivity to DNA Damage Agents

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MIA PaCa-2 and HeLa cells were seeded at 5 × 10⁵ cells/60 mM dish. U2OS cells were seeded at either 5 × 10⁵ cells/60 mM dish, or 4 × 10⁴ cells/60 mM dish. Each dish was then transfected with either 800 ng/expression plasmid (FANCA-plvx IRES-Neo, FANCG-plvx-IRES-Neo, FAAP20-pcDNA, plvx-IRES-Neo empty vector) or 80 pmol targeting siRNA (FANCA, FANCG, FAAP20 Smartpool-Dharmacon), or scrambled siCtrl (ON-TARGETplus Non-targeting Control Pool-Dharmacon). In all, 3 μl Lipofectamine 2000/expression plasmid or 4 μl RNAimax/siRNA target was diluted in Optimem reduced serum-medium (Gibco), and added to indicated nucleic acids, and incubated at RT for 25 min. Transfection complexes were then added to cell dishes overnight, and media was changed 16 h later. Twenty-four hours later, cells were harvested, counted, and seeded into 6-well plates at 500 cells/well. Twenty-four hours after seeding wells, plates were either treated with 5 μM DI03 (Sigma), 10 uM AZD2461 (Sigma), 0.5 Gy IR, or left untreated. Plated cells grew for 14 days before staining with crystal violet.

Palovcak A., Yuan F., Verdun R., Luo L, & Zhang Y. (2023). Fanconi anemia associated protein 20 (FAAP20) plays an essential role in homology-directed repair of DNA double-strand breaks. Communications Biology, 6, 873.

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Mutant p53 Radiosensitization in HCT116

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HCT116 p53-/-were plated in 6-well plates at a density of 2 × 10 5 cells/well in 2 ml and, the day after, transfected with empty vector or pcDNA3-p53R273H vector for mutant p53 expression, by using Lipofectamine 3000 (Invitrogen) according to the manufacturer's instructions. After 24 h of transfection, cells were pre-treated with AZD2461 (30 μM) (Sigma Aldrich) for 5 h, then irradiated with 1 Grey (1Gy) radiation and cultured for the next 24 h. Untreated cells were used as control.

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Cell Proliferation Evaluation by MTT Assay

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Cell proliferation was evaluated by MTT assay (Sigma Aldrich). 5 × 10 3 cells/well were plated in 96-well plates in 100 μL of complete medium. The day after, cells were treated with different doses of AZD2461 (30 μM, 60 μM, 120 μM and 240 μM) for 72 h. In some experiments 5 × 10 3 were plated in 96-well plates in 100 μL of complete medium and transfected as previously described or left un-transfected. After 24 h cells were pre-treated with AZD2461 (30 μM) (Sigma Aldrich) for 5 h, then irradiated with 1 Grey (1Gy) radiation and cultured for the next 72 h. Untreated cells were used as control. MTT assay was performed following manufacturer's instruction. The plates were analyzed by VICTOR Multilabel Plate Reader (PerkinElmer). The experiments were performed in triplicate and repeated three times.

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