N ethyl n nitrosourea

Manufactured by Merck Group

Sourced in United States

N-ethyl-N-nitrosourea is a chemical compound that serves as a laboratory reagent. It is commonly used as a mutagen in scientific research to induce genetic alterations in experimental organisms. The core function of N-ethyl-N-nitrosourea is to facilitate the study of genetic processes and the effects of mutagenic agents on living systems.

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Lab products found in correlation

Low melting point agarose, by Thermo Fisher Scientific (1 mentions) Ethyl methanesulfonate, by Merck Group (1 mentions) Triton x 100, by Thermo Fisher Scientific (1 mentions) Countbright absolute count beads and fetal bovine serum, by Thermo Fisher Scientific (1 mentions) Tris hydroxymethyl aminomethane, by Serva Electrophoresis (1 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Sodium hydroxide (naoh), by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Quadromacs separator, by Miltenyi Biotec (1 mentions) Inositol hexaphosphate, by Merck Group (1 mentions) P akt, by Santa Cruz Biotechnology (1 mentions) P stat3, by Santa Cruz Biotechnology (1 mentions) Caspase 3, by Santa Cruz Biotechnology (1 mentions) Stat3, by Santa Cruz Biotechnology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Cyclin d1, by Santa Cruz Biotechnology (1 mentions) Caspase 9, by Santa Cruz Biotechnology (1 mentions) Rneasy minielute cleanup kit, by Qiagen (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

N-ethyl-N-nitrosourea (ENU)

8 protocols using n ethyl n nitrosourea

Developing ATB-Resistant Cancer Cell Lines

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HCT116 and LOVO cells (1 × 10⁷) were exposed to 100 mg/mL N-ethyl-N-nitrosourea (Sigma-Aldrich, Burlington, MA, USA) for 24 hours, followed by treatment with a gradient concentration of ATB to induce ATB resistance. During the first five days, HCT116 and LOVO cells were exposed to ATB (4 μg/mL), and the medium was changed every day. In the next two months, 6, 8, 10, and 12 μg/mL of ATB were used to treat the cells [32 (link)]. Finally, about 100 variable cells exposed to ATB (12 μg/mL) were recovered. After proliferation for about one month, the ATB-resistant HCT116 and LOVO cells, named HCT116/ATB and LOVO/ATB separately, were used for functional assays.

Pan D., Chen K., Chen P., Liu Y., Wu Y, & Huang J. (2023). Downregulation of LncRNA SNHG7 Sensitizes Colorectal Cancer Cells to Resist Anlotinib by Regulating miR-181a-5p/GATA6. Gastroenterology Research and Practice, 2023, 6973723.

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ENU Mutagenesis Screening for Lethal Rescues

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ENU mutagenesis was performed as previously described [5 (link)], with all mice on the C57BL/6J genetic background. Briefly, 189 F5^L/L male mice (6–8 weeks old) were administered three weekly intraperitoneal injections of 90 mg/kg of ENU (N-ethyl-N-nitrosourea, Sigma-Aldrich). Eight weeks later, 182 surviving males were mated to F5^L/+Tfpi^+/- females and their G1 progeny were genotyped at age 2–3 weeks to identify viable F5^L/LTfpi^+/- offspring (‘rescues’). F5^L/LTfpi^+/- G1 rescues were crossed to F5^L/L mice on the C57BL/6J genetic background (backcrossed >20 generations) and transmission was considered positive with the presence of one or more rescue progeny. Theoretical mapping power in rescue pedigrees was estimated by 10,000 simulations using SIMLINK software [36 ].

Tomberg K., Westrick R.J., Kotnik E.N., Cleuren A.C., Siemieniak D.R., Zhu G., Saunders T.L, & Ginsburg D. (2018). Whole exome sequencing of ENU-induced thrombosis modifier mutations in the mouse. PLoS Genetics, 14(9), e1007658.

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Mutagenicity Assay Protocol

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Acetaminophen (CAS number 103‐90‐2) was obtained from Specgx, Raleigh, NC. N‐ethyl‐N‐nitrosourea (ENU; CAS number 759‐73‐9) and ethyl methanesulfonate (EMS; CAS number 62‐50‐0) were purchased from Sigma Aldrich, St. Louis, MO. Hydroxypropyl methylcellulose (Methocel F4M Premium) was obtained from Dow Chemical Company, Midland, MI.
All reagents to perform the micronucleus assay were from Rat MicroFlow® PLUS kits (Litron Laboratories, Rochester, NY). Most reagents to perform the Pig‐a assay were from prototype Rat MutaFlow® kits (Litron Laboratories). Additional supplies included Lympholyte®‐Mammal cell separation agent from CedarLane, Burlington, NC; Anti‐PE MicroBeads, LS Columns and a QuadroMACS™ separator from Miltenyi Biotec, Bergisch Gladbach, Germany; and CountBright™ Absolute Count Beads and fetal bovine serum from Invitrogen, Carlsbad, CA. Reagent supplies to perform the comet assay included normal melting point agarose, ethylenediaminetetraacetic acid disodium salt dihydrate (EDTA‐Na₂.2H₂O), sodium chloride (NaCl), and sodium hydroxide (NaOH) from Sigma Aldrich; dimethyl sulfoxide (DMSO) and ethanol from Merck, Darmstadt, Germany; Hank's balanced salt solution and Ca2+/Mg2+ free phosphate buffered saline (PBS) and Triton X‐100 from ThermoFisher, Waltham, MA; low melting point agarose from Invitrogen; and Tris hydroxymethyl aminomethane from Serva, Heidelberg, Germany.

van der Leede B., Weiner S., Van Doninck T., De Vlieger K., Schuermans A., Tekle F., Geys H., van Heerden M., De Jonghe S, & Van Gompel J. (2020). Testing of acetaminophen in support of the international multilaboratory in vivo rat Pig‐a assay validation trial. Environmental and Molecular Mutagenesis, 61(5), 508-525.

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Induced Mutagenesis in Arabidopsis

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PBG seeds (0.2 g) were hydrated in 45 ml of ddH₂O with 0.005% Tween-20 and left on a tube rotator for 4 h. The seeds were washed with ddH₂O twice and then soaked in 1 mM N-ethyl-N-nitrosourea or 0.2% ethyl methanesulfonate (Sigma-Aldrich, St. Louis, MO) solutions for 15 h with rotation. Then, the seeds were washed with ddH₂O eight times, stratified in the dark at 4 °C for 4 days, and then sown onto large plates. The seedlings (M1 generation) were transferred to soil, and the progeny (M₂ generation) were collected from individual plants.

Yoo C.Y., Pasoreck E.K., Wang H., Cao J., Blaha G.M., Weigel D, & Chen M. (2019). Phytochrome activates the plastid-encoded RNA polymerase for chloroplast biogenesis via nucleus-to-plastid signaling. Nature Communications, 10, 2629.

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Leukemic Mice Treated with PF-0077736

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Experiments involving mice were performed in agreement with Italian guidelines and after the approval of the Institutional Review Board of the European Institute of Oncology. Leukemic mice were generated by a single administration of the tumorigenic agent n-ethyl-n-nitrosourea (Sigma, 50 mg/kg intraperitoneally). Spleen cells from leukemic C57BL6/Ly5.2 mice were injected intravenously (2 × 10⁶ cells/mouse) into non-irradiated, recipient C57BL6/Ly5.1 mice. PF-0077736 was administered intraperitoneally starting from day 3 after the transplant of leukemic cells. Mice received doses of PF-0077736 (40 mg/kg each dose) every 3 days for four treatments (q3dx4). Kaplan-Meyer rank test was used to compare the survival rate between treated and control mice.

Iacobucci I., Di Rorà A.G., Falzacappa M.V., Agostinelli C., Derenzini E., Ferrari A., Papayannidis C., Lonetti A., Righi S., Imbrogno E., Pomella S., Venturi C., Guadagnuolo V., Cattina F., Ottaviani E., Abbenante M.C., Vitale A., Elia L., Russo D., Zinzani P.L., Pileri S., Pelicci P.G, & Martinelli G. (2015). In vitro and in vivo single-agent efficacy of checkpoint kinase inhibition in acute lymphoblastic leukemia. Journal of Hematology & Oncology, 8, 125.

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Anticarcinogenic Potential Evaluation

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N-ethyl-N-nitrosourea and inositol hexaphosphate were procured from Sigma Co. (USA). [methyl-³H] thymidine (specific activity 17.20 Ci/mmol) was provided by BRIT, Mumbai, India. caspase-3 or -9 enzyme activity assay kit was procured from R&D system, USA. The Super Signal West Femto Max Substrate was procured from Thermo Fischer Scientific, USA. Primary antibodies for PCNA, cyclin D1, NF-κB (p50), IL-6, STAT3, pSTAT3, pAkt, PARP, caspase-3 and caspase-9 were purchased from Santa Cruz Biotechnologies, USA and antibodies for COX-2, Akt and β-actin were from Cell Signalling Technology, USA. The rest of the chemicals were purchased from local commercial sources and were of analytical grade.

Sahay S., Tiwari P, & Gupta K.P. (2015). Onset of the lymphocytic infiltration and hyperplasia preceding the proliferation in F1 mouse lungs from the N-ethyl-N-nitrosourea exposed mothers: Prevention during the lactation period by inositol hexaphosphate. Toxicology Reports, 2, 590-599.

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ENU-Induced Mutagenesis of EGFR Cell Lines

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N-ethyl-N-nitrosourea (ENU) was purchased from Sigma Aldrich and mutagenesis was carried out as previously described (24 (link)). Briefly, L858R, L858R/T790M, DelE746_A750 and DelE746_A750 Ba/F3 cells (1 X 10⁶ cells/ml) were exposed to ENU (50 μg/ml) for 24 hours. Cells were then washed 3 times with RPMI, and expanded in growth media for 5–7 days. Cells were subsequently cultured in 96-well plates (1 X 10⁴ cells/well; total 5 X 10⁵ cells per inhibitor) in the presence of 100 nM WZ4002, 1 μM WZ4002. Wells were observed for growth by visual inspection and resistant wells were expanded in the presence of the corresponding inhibitor. Total RNA was isolated from drug resistant cell lines using Trizol^™ (Invitrogen, Carlsbad, CA) and purified using RNeasy^™ minielute cleanup kit (Qiagen, Valencia, CA). cDNA was transcribed from 2μg of total RNA with Superscript II Reverse Transcriptase (Invitrogen Life technologies, Carlsbad, CA). The cDNA was used as template for subsequent sequencing of the EGFR tyrosine kinase domain (exons 18–21).

Ercan D., Choi H.G., Yun C.H., Capelletti M., Xie T., Eck M.J., Gray N.S, & Jänne P.A. (2015). EGFR mutations and resistance to Irreversible pyrimidine based EGFR inhibitors. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(17), 3913-3923.

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Icaritin and ENU Exposure Protocol

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Cayman Chemical supplied the Icaritin (ICT; cat. no. Cay20236-500, Purity > 98%), and Sigma Aldrich supplied the N-ethyl-N’-nitrosourea (ENU; St. Louis, MO, USA).

Hou X., Han Y., Hirad A.H., Alarfaj A.A, & Liu L. (2024). N-Ethyl-N-Nitrosourea Induced Leukaemia in a Mouse Model: Protective Effect of Icaritin via Inhibition of IL-6/JAK2/STAT3 Pathway Causes Apoptosis. Journal of Inflammation Research, 17, 777-790.

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