Ethyl methanesulfonate ems

Manufactured by Merck Group

Sourced in United States

Ethyl methanesulfonate (EMS) is a chemical compound used in various laboratory applications. It functions as a mutagenic agent, capable of inducing genetic mutations in biological systems. The core purpose of EMS is to facilitate the study of genetic processes and the evaluation of the effects of chemical compounds on genetic material.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Methyl methanesulfonate mms, by Merck Group (1 mentions) Cometassay, by Bio-Techne (1 mentions) Comet analysis software, by Bio-Techne (1 mentions) Evos m7000 imaging system, by Thermo Fisher Scientific (1 mentions)

7 protocols using ethyl methanesulfonate ems

Genotoxicity Evaluation with EMS

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All chemicals were purchased from Sigma-Aldrich, unless differently specified in the text. Ethyl methanesulfonate (EMS) was purchased from Sigma-Aldrich, Ireland, and used according to manufacturer’s specification. EMS was diluted in cell culture medium and used at a concentration of 15 mM as the genotoxic positive control for experiments.

Tutty M.A., Vella G., Vennemann A., Wiemann M, & Prina-Mello A. (2022). Evaluating nanobiomaterial-induced DNA strand breaks using the alkaline comet assay. Drug Delivery and Translational Research, 12(9), 2243-2258.

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Subacute In Vivo Genotoxicity Assay Protocol

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The subacute combined peripheral blood erythrocyte micronucleus (MN) and alkaline pH>13 comet assay was conducted at Integrated Laboratory Systems, Inc. (Durham, NC) (Recio et al. 2010 (link); Witt et al. 2010 (link)). Tissues sampled in the comet assay included forestomach, liver, colon, and blood leukocytes. The MN test measures chromosomal damage in the form of structural and/or numerical changes, while the alkaline comet assay measures primary DNA damage in the form of strand breaks, adducts, and alkali labile sites. Briefly, male B6C3F1/N mice (Charles River Laboratories, Portage, MI, 8–16 wk of age) were exposed to the vehicle control (corn oil), MNA (500, 1000, 1500 or 2000 mg MNA/kg/day), or the positive control (100 mg/kg/day ethyl methanesulfonate, EMS, Sigma-Aldrich) daily for 4 days by oral gavage (N=5). The dose limit for both assays, in the absence of overt toxicity, is 2000 mg/kg/day (OECD 489 2016 ; OECD 2016 ). Four hours after the final treatment with MNA, the mice were euthanized via carbon dioxide asphyxiation. Slides from peripheral blood and bone marrow were immediately prepared, fixed, and stained with acridine orange for MN analysis; tissue samples for the comet assay were collected, processed, flash frozen, and stored in a −80°C freezer until thawed for analysis, as previously described (Recio et al. 2010 (link); Witt et al. 2010 (link)).

Frawley R.P., Witt K.L., Cunny H., Germolec D.R., Jackson-Humbles D., Malarkey D., Shockley K.R., Stout M., Travlos G., Buccellato M., Fallacara D., Harris S., Kissling G.E., Manheng W., Surh I.I., White K J.r, & Auerbach S.S. (2020). Evaluation of 2-methoxy-4-nitroaniline (MNA) in hypersensitivity, 14-day subacute, reproductive, and genotoxicity studies. Toxicology, 441, 152474.

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Phytochemical Profiling of Aloe ferox Juice

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Dried Aloe ferox juice was kindly supplied by Erbofrut s.n.c., Roccavione (Italy). The characterization of dried Aloe ferox juice (provided by Indena, SpA, Milan, Italy) is reported in Table 1. Anthraquinones were identified by ultra-high-performance liquid chromatography, chromones by nuclear magnetic resonance analysis, and mono- and disaccharides (fructose, galactose, glucose, fructose, lactose, maltose) by using a gas chromatographic method after derivatization. A Karl Fisher system was used to detect water, and total amino acids were determined by high-performance liquid chromatography. Metals were assayed using inductively coupled plasma mass spectrometry, while inorganic and organic anions were assayed by ionic chromatography.Table 1

Composition of dried Aloe ferox juice.

Table 1

Antraquinones	%	Chromones	%	Other analytes	%
Aloin (A+B)	8.30	Aloesin	28.70	Glucose	0.30
Aloe-emodin	0.20	Aloeresin	32.20	Water	6.70
Rhein	0.03			Total amino acids	0.55
Emodin	0.01			Metals	0.12
Chrysophanol	0.01			Inorganic anions	0.05
Physcion	n.d.			Organic ions	0.27

The % represents w/w. The sum is 77.44 %.

Ethyl methanesulfonate (EMS, Sigma) was used as a positive control and prepared in water.

Galli C.L., Cinelli S., Ciliutti P., Melzi G, & Marinovich M. (2021). Lack of in vivo genotoxic effect of dried whole Aloe ferox juice. Toxicology Reports, 8, 1471-1474.

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Chlorophyll and Mutagen Dose Preparation

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2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx, C₁₁H₁₁N₅, CAS No. 77500-04-0) was purchased from Toronto Research Chemicals; chlorophyll a (C₅₅H₇₂N₄O5_Mg, CAS No. 479-61-8), chlorophyll b (C₅₅H₇₀MgN₄O₆, CAS No. 519-62-0), dimethyl sulfoxide (DMSO, CAS No. 67-68-5), ethyl methane sulfonate (EMS; CAS No. 62-50-0) were purchased from Sigma. Chlorophyll a and b were dissolved in DMSO at 0.5% and diluted in distilled water to obtain the test concentrations (0.5, and 1 µM). Mutagen was dissolved in DMSO at 0.5% and diluted in distilled water to obtain the test concentrations (4.69, 9.38 and 23.45 µM).

Kocaoğlu Cenkci S, & Kaya B. (2022). Effects of Chlorophyll a and b in Reducing Genotoxicity of 2-Amino-3,8-dimethylimidazo[4,5-F]quinoxaline (MeIQx). Biology, 11(4), 602.

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In Vivo Genotoxicity Assessment of CGTase and Sodium Sulfate

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GLP studies were conducted in compliance with OECD TG 474 [19] and in general accordance with OECD TG 489 [20] . Dose range finding studies were conducted to identify appropriate dose levels. Male and female mice were administered CGTase or sodium sulfate via oral gavage up to and including dose levels of 2000 mg/kg/day (test guideline limit dose) for three consecutive days. No marked changes in body weights were measured nor adverse clinical observations found in mice administered either chemical and there were no abnormal gross necropsy organ observations. Therefore, for the definitive studies, male and female B6C3F1 mice (5 animals/dose group) were administered CGTase or sodium sulfate at 1000, 1500, or 2000 mg/kg/day, vehicle (deionized water), or the positive control compound, ethyl methanesulfonate (EMS) (Sigma-Aldrich, St. Louis, MO) in 0.9% saline (Ricca Chemical Company, Arlington, TX) at 150 mg/kg/day, daily for three days by oral gavage. Three hours after the final dose, peripheral blood was collected for flow cytometric analysis of MN, and liver, duodenum, and stomach tissues were collected, frozen in liquid nitrogen, and stored at ‐80 °C until analysis by the comet assay [24] (link).

Maronpot R.R., Hobbs C.A., Davis J., Swartz C., Boyle M., Koyanagi M, & Hayashi S.M. (2016). Genetic and rat toxicity studies of cyclodextrin glucanotransferase. Toxicology Reports, 3, 381-392.

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Dose-dependent Cytotoxic Effects of Alkylating Agents

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Alkylating agents, methyl methanesulfonate (MMS) (CAS number 66-27-3), and ethyl methanesulfonate (EMS) (CAS number 62-50-0) were obtained from Sigma Co., St. Louis, MO, USA, and colchicine (CAS number 64-86-8) was obtained from Himedia, Pvt. Ltd., Mumbai, India. Giemsa stain and other chemicals of analytical grade were commercially available. EMS and MMS were dissolved in 0.9% NaCl to obtain the required concentrations. Freshly prepared solutions of these agents were used each time. The conditioning and challenging doses of EMS (80 and 240 mg/kg body weight) and MMS (50 and 150 mg/kg body weight) were selected from the earlier experiments [28 (link), 29 (link)]. Even though these were selected from treated normal animals, the effect of these along with the range (namely, 25–160 mg/kg body weight of MMS and 50–300 mg/kg body weight of EMS) of doses was analyzed in EAC cells by preliminary pilot toxicity studies.

Mahadimane P.V, & Vasudev V. (2014). Inducible Protective Processes in Animal Systems XIII: Comparative Analysis of Induction of Adaptive Response by EMS and MMS in Ehrlich Ascites Carcinoma Cells. Scientifica, 2014, 703136.

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Alkaline Comet Assay for DNA Damage

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To assess SSB, SKOV3, and COV362, cells were exposed to NE at the timepoints required for each experiment: 1 h or 24 h. After NE exposure, the cells were collected and processed to perform the alkaline version of the CometAssay^® as described by the manufacturer (Trevigen, Gaithersburg, MD, USA). The nuclei were stained with Yoyo-1^® and visualized using the EVOS™ M7000 Imaging System (Thermo Scientific, Rockford, IL, USA). Fifty nuclei were randomly selected and analyzed for percentage of DNA in tail using the Comet Analysis Software (Trevigen, Gaithersburg, MD, USA). Ethyl methanesulfonate (EMS) (Sigma, St. Louis, MO, USA) was used as a positive control at a concentration of 12 mM for 4 h.

Lamboy-Caraballo R., Ortiz-Sanchez C., Acevedo-Santiago A., Matta J., N.A. Monteiro A, & N. Armaiz-Pena G. (2020). Norepinephrine-Induced DNA Damage in Ovarian Cancer Cells. International Journal of Molecular Sciences, 21(6), 2250.

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