Ecs5000l

Manufactured by Euroclone

The ECS5000L is a laboratory equipment designed for scientific research and analysis. It is a high-performance instrument that provides accurate and reliable measurements. The core function of the ECS5000L is to perform electrochemical analysis, such as cyclic voltammetry, potentiometry, and electrochemical impedance spectroscopy. The specific details and intended use of this product are not included in this description.

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Lab products found in correlation

Ecb3001d, by Merck Group (2 mentions) U3003, by Merck Group (2 mentions) Ecb3000d, by Euroclone (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Lipofectamin 2000, by Thermo Fisher Scientific (1 mentions) Effectene, by Qiagen (1 mentions) Fetal bovine serum (fbs), by Capricorn (1 mentions) Penicillin, by Euroclone (1 mentions) Streptomycin, by Euroclone (1 mentions) Dmem f12, by Euroclone (1 mentions) 96 wells plate, by Corning (1 mentions) Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions) Poly d lysine, by Merck Group (1 mentions) Torin 1, by MedChemExpress (1 mentions) Evos m5000 imaging system, by Thermo Fisher Scientific (1 mentions) G7021, by Merck Group (1 mentions) Ovsaho, by American Type Culture Collection (1 mentions) Ecb7501l, by Euroclone (1 mentions)

4 protocols using ecs5000l

Lung Cancer Cell Line H1299 Culture

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The non-small cell lung cancer cell line H1299 (ATCC) was used because of its p53 deletion and non-detectable or low endogenous levels of p63 and p73, respectively, resulting in low background levels for the functional assays in this study. H1299 cells were cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% Fetal Bovine Serum (FBS, Capricorn Scientific), 100 U/ml penicillin and 100 µg/ml streptomycin (Thermo Fisher Scientific). HEK293T cells (a gift from Gian-Paolo Dotto) were used for virus production and cultured in Dulbecco’s in Dulbecco Modified Eagle Medium (Sigma) supplemented with 10% FBS (#ECS5000L, Euroclone) and 2 mM Glutamine. Newborn human dermal fibroblasts (HDF, #C0045C, Thermo Fisher Scientific) were cultured in DMEM high glucose with the 10% FBS (#ECS5000L, Euroclone). BJ-HDF were cultured in DMEM-F12 (#ECM0090L, Euroclone) supplemented with either 10% FBS (#ECS5000L, Euroclone) or 10% tetracycline negative FBS (#ECS0182L, Euroclone). All cell lines were cultured at 37 °C and 5% CO₂ and were routinely tested for mycoplasma contaminations.
H1299 cells were transfected using either the Effectene (Qiagen) or the Lipofectamin 2000 (Thermo Fisher Scientific) transfection reagent according to the manufacturers’ protocols.

Osterburg C., Ferniani M., Antonini D., Frombach A.S., D’Auria L., Osterburg S., Lotz R., Löhr F., Kehrloesser S., Zhou H., Missero C, & Dötsch V. (2023). Disease-related p63 DBD mutations impair DNA binding by distinct mechanisms and varying degree. Cell Death & Disease, 14(4), 274.

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Cell Culture Techniques for Functional Analyses

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HeLa cells were grown in DMEM (Euroclone) cell culture medium, while CFPAC-1 cells were grown in DMEM-F12 (Euroclone). Cell culture media were supplemented with 10% FBS (ECS5000L Euroclone), 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin (Euroclone). Primary fibroblasts were grown in RPMI medium (Euroclone) supplemented with 20% FBS (ECS5000L Euroclone), 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin (Euroclone). For sphingolipid transport functional analysis, for autophagic vacuoles detection, and for DENND5B immunolocalization studies, 30 × 10⁵ per well primary fibroblasts derived from affected individuals’ dermal biopsies, HeLa, or CFPAC-1 cells were plated in 96-wells plates (Corning). For CFPAC-1 cells, plates were coated with poly-D-lysine (Merck KGaA) to increase cellular adherence to the bottom of the wells. Cells were imaged 24 h after plating.

Scala M., Tomati V., Ferla M., Lena M., Cohen J.S., Fatemi A., Brokamp E., Bican A., Phillips JA I.I.I., Koziura M.E., Nicouleau M., Rio M., Siquier K., Boddaert N., Musante I., Tamburro S., Baldassari S., Iacomino M., Scudieri P., Rosenfeld J.A., Bellus G., Reed S., Al Saif H., Russo R.S., Walsh M.B., Cantagrel V., Crunk A., Gustincich S., Ruggiero S.M., Fitzgerald M.P., Helbig I., Striano P., Severino M., Salpietro V., Pedemonte N, & Zara F. (2024). De novo variants in DENND5B cause a neurodevelopmental disorder. American Journal of Human Genetics, 111(3), 529-543.

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Fibroblasts from CMT2A Patient

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Primary fibroblasts from a young patient affected by CMT2A^MFN2 (c.650G>T/p.Cys217Phe) and a healthy control (individual with no histological or biochemical signs of mitochondrial disease), were obtained as reported in ^{8 (link)} after informed consent. Cells were grown in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; EuroClone, ECB7501LX10) supplemented with 10% (v/v) fetal bovine serum (FBS; EuroClone, ECS5000L), 1% (v/v) L-glutamine (E EuroClone, ECB3000D), 1% (v/v) penicillin/streptomycin (EuroClone, ECB3001D), 50μg/ml of uridine (Sigma-Aldrich, U3003), in a humidified incubator at 37°C and 5% CO2 avoiding confluence at any time. All experiments were performed on cells with similar passage numbers, ranging from 5 to 8, to avoid any artefact due to senescence. For the experiments, growing cells were plated on sterile plastic dishes or flasks and allowed to adhere for at least 24 h before use. Torin1 (MedChemExpress, USA) was used at 0.1, 0.25 and 0.5 μM for 72 h, and DMSO as a vehicle.

Zanfardino P., Amati A., Petracca E.A., Santorelli F.M, & Petruzzella V. (2022). Torin1 restores proliferation rate in Charcot-Marie-Tooth disease type 2A cells harbouring MFN2 (mitofusin 2) mutation. Acta Myologica, 41(4), 201-206.

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Ovarian Cancer Cell Line Cultivation and Treatment

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The human ovarian cancer cell lines SKOV3 and OVSAHO were purchased from ATCC^® (Manassas, VA, USA) and JCRB Cell Bank (Japan), respectively. Cells were cultured in High Glucose Dulbecco's Modified Eagle's Medium (DMEM) with sodium pyruvate (Euroclone #ECB7501L) supplemented with 10% FBS South America origin EU Approved (Euroclone #ECS5000L), 2 mM L-Glutamine (Euroclone #ECB3000D), 1% penicillin/streptomycin (Euroclone #ECB3001D) and 50 µg/mL uridine (Sigma–Aldrich #U3003) and maintained at 37 °C in a humidified atmosphere with 5% CO₂. For high- or low-glucose experiments, cells were grown for 24 hours in DMEM (Gibco #11966025) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin, 50 µg/mL uridine, 1 mM sodium pyruvate (Sigma–Aldrich #P2256) and 25 mM or 5 mM D-(+)-glucose (Sigma Aldrich #G7021). Where indicated, cells were treated with 1 µM N4-[2-(4-Phenoxyphenyl) ethyl]-4,6-quinazolinediamine [EVP-4593; Sigma–Aldrich #SML0579] at multiple timepoints (6 h, 12 h, 24 h) and compared to Time 0 (T0). EVOS M5000 Imaging System (Thermo Fisher Scientific #AMF5000) was used for cell line monitoring.

De Luise M., Sollazzo M., Lama E., Coadă C.A., Bressi L., Iorio M., Cavina B., D’Angelo L., Milioni S., Marchio L., Miglietta S., Coluccelli S., Tedesco G., Ghelli A., Lemma S., Perrone A.M., Kurelac I., Iommarini L., Porcelli A.M, & Gasparre G. (2022). Inducing respiratory complex I impairment elicits an increase in PGC1α in ovarian cancer. Scientific Reports, 12, 8020.

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