Neomycin

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, China, Spain, Macao, Belgium

Neomycin is an antibiotic medication produced by the Merck Group. It is a type of aminoglycoside antibiotic that functions by inhibiting protein synthesis in bacterial cells.

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Lab products found in correlation

Ampicillin, by Merck Group (148 mentions) Vancomycin, by Merck Group (120 mentions) Metronidazole, by Merck Group (109 mentions) Streptomycin, by Merck Group (69 mentions) Penicillin, by Merck Group (48 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (37 mentions) Gentamicin, by Merck Group (30 mentions) Puromycin, by Merck Group (25 mentions) Polymyxin b, by Merck Group (24 mentions) Kanamycin, by Merck Group (18 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (14 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (13 mentions) Dimethyl sulfoxide (dmso), by Merck Group (12 mentions) Dmem f12, by Thermo Fisher Scientific (12 mentions) Amphotericin b, by Merck Group (11 mentions) Chloramphenicol, by Merck Group (11 mentions) Vancomycin, by MP Biomedicals (10 mentions) Bacitracin, by Merck Group (10 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (10 mentions) Streptomycin, by Thermo Fisher Scientific (10 mentions)

540 protocols using neomycin

Astaxanthin Protects Utricles from Neomycin

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Utricles were divided into five groups as follows: control, neomycin (2 mM, Sigma Chemical Co.), neomycin+astaxanthin (1 µM), neomycin+astaxanthin (10 µM), and neomycin+astaxanthin (1–100 µM). In the control group, only the vehicle was added to the medium. In the neomycin group, neomycin was added to the culture at a concentration of 2.0 mM. In the neomycin+astaxanthin groups, in addition to 2 mM neomycin, astaxanthin was added at 1 µM, 10 µM, and 100 µM, respectively. The utricles were incubated for 1 h before exposure to neomycin.
All free-floating utricles were incubated for 24 h at 37 °C in 24-well tissue culture plates in a 5% CO₂ and 95% air environment. After all culture protocols, utricles were fixed with 4% paraformaldehyde (PFA) for 1 h at room temperature. Otoconia from the fixed utricles were gently removed using a stream of phosphate-buffered saline (PBS) applied through a syringe and a 28G needle. After the samples were rinsed with PBS, they were subsequently used in assays.

Kobayashi Y., Sugahara K., Takemoto Y., Tsuda J., Hirose Y., Hashimoto M, & Yamashita H. (2023). Protective effect of astaxanthin nanoemulsion on mammalian inner ear hair cells. PeerJ, 11, e15562.

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Genetic Manipulation of Colon Cancer Cells

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The PLD1 shRNA vectors were generated by ligation of vector pCMV-RFP. To establish shRFP, shPLD1-a, and shPLD1-b stable cell lines, xenografted DLD1, HCT116, and SW480 cells were transfected using control or two types of shRNA for PLD1 for 72 h under sphere cultured conditions, and then cells were selected with 500 µg/ml neomycin (Sigma-Aldrich) for 14 d. To avoid clonal variation, the stable cell lines were established from the mixed population of multiple neomycin-resistant clones. RFP-positive cells were sorted by using flow cytometry (FACS). shPLD1-a plus shE2F1, anti-EV, or anti–miR-4496 stable cell lines were established by transduction of their lentiviruses with 6 µg/ml polybrene (Sigma-Aldrich) and 500 µg/ml neomycin/4 µg/ml puromycin double selection.

Kang D.W., Choi C.Y., Cho Y.H., Tian H., Di Paolo G., Choi K.Y, & Min D.S. (2015). Targeting phospholipase D1 attenuates intestinal tumorigenesis by controlling β-catenin signaling in cancer-initiating cells. The Journal of Experimental Medicine, 212(8), 1219-1237.

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Establishment of Stable Cell Lines Expressing KIF1C

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Stable cell lines were cultured under standard conditions in Dulbecco's Modified Eagle's Medium (DMEM) high glucose (Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific) and 1% neomycin (Sigma-Aldrich) for HEK 293 cells, and in DMEM/F12 (Sigma-Aldrich) with 15% FBS and 1% neomycin for SH-SY5Ys. Cells were trypsinized when confluent and plated according to the required cell number. To generate cell lines stably expressing KIF1C, we expressed plasmids containing the HA-tagged wild-type (WT) or mutant (mt) (c.305G > C; p.G102A [Caballero Oteyza et al. 2014 (link)]) KIF1C open reading frame in a pSF-CMV-UB-NEO/G418 ASCI (Sigma-Aldrich) backbone and cultured cells under neomycin selection conditions. Control cell lines were generated with a plasmid expressing WT HA-tagged KIF5A, a gene mutated in Hereditary Spastic Paraplegia Type SPG10, and a plasmid only expressing the HA-tag.
To induce neuronal differentiation of SH-SY5Y cells, the medium of 60% confluent cells was switched to DMEM/F12 with reduced serum concentration (5% FBS), 1% neomycin and addition of 10 µM retinoic acid (RA) (StemCell Technologies). At day 6 the cells were washed twice with warm PBS (Sigma-Aldrich) and the media was changed to serum-free DMEM/F12 with 1% neomycin and 25 ng/mL BDNF (PeproTech). The cells were used from day 12 to day 18 after differentiation.

Nagel M., Noß M., Xu J., Horn N., Ueffing M., Boldt K, & Schüle R. (2023). The kinesin motor KIF1C is a putative transporter of the exon junction complex in neuronal cells. RNA, 29(1), 55-68.

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Modeling Ototoxicity, Noise, and LPS-Induced Hair Cell Injury

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To construct the ototoxic drug-induced HC injury model in the neonatal stage, cochlear epithelia samples were obtained for explant culture from P2 transgenic mice that had received tamoxifen (Sigma, T5648-5G) administration at P 0–1. The tissues were treated with 1 mM neomycin (Sigma) for 6 h and then harvested after an additional 24 h in the absence of neomycin.
To construct the noise-induced HC injury model, the hearing function of P30 mice that had received tamoxifen administration at P21–P22 were examined by ABR test 24 h before noise exposure. The mice were then exposed to broadband noise at 116 dB for 2 h. Hearing function was evaluated again by ABR test 7 days after noise exposure, and the mice were killed and cochlear samples were harvested for morphological analysis.
To construct the lipopolysaccharide (LPS)-induced HC injury model, mice were given 10 μl LPS (Sigma-Aldrich, L2880) at 1, 2, or 4 mg/kg by tympanum injection and killed 3 days later.

Chen D., Jia G., Zhang Y., Mao H., Zhao L., Li W., Chen Y, & Ni Y. (2022). Sox2 overexpression alleviates noise-induced hearing loss by inhibiting inflammation-related hair cell apoptosis. Journal of Neuroinflammation, 19, 59.

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Cochlear Sensory Epithelium Neomycin Exposure

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Cochlear sensory epithelium was dissected from postnatal day (P)3 wild-type FVB mice and cultured in DMEM/F12 (Gibco, Gaithersburg, MD, USA, 11330-032) supplemented with 2% B27 (Invitrogen, Waltham, MA, USA, 17504044), 1% N-2 (Invitrogen, Waltham, MA, USA, 17502-048), and 50 μg/ml ampicillin (Sigma–Aldrich, St. Louis, MO, USA, P0781). In the experimental group, the cochleae were treated with 0.5 mM neomycin (Sigma–Aldrich, St. Louis, MO, USA, N6386-5G) and 1 μM blebbistatin (dissolved in DMSO, Boehringer Ingelheim Pharma GmbH, Biberach an der Riß, Germany) for 12 h and allowed to recover for another 12 h. Equivalent amounts of DMSO (Sigma–Aldrich, St. Louis, MO, USA, D8371) were added to the control and neomycin-only groups. The tissues were cultured at 37°C with 5% CO₂.

Gao S., Cheng C., Wang M., Jiang P., Zhang L., Wang Y., Wu H., Zeng X., Wang H., Gao X., Ma Y, & Chai R. (2020). Blebbistatin Inhibits Neomycin-Induced Apoptosis in Hair Cell-Like HEI-OC-1 Cells and in Cochlear Hair Cells. Frontiers in Cellular Neuroscience, 13, 590.

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CRISPR-Mediated Genetic Modification in ES Cells

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E14 and HM1 ES cells were cultured on gelatin-coated plates with standard ES cell culture conditions.
Before transfection the cells were seeded into 6 well plates (approximately 5 × 10⁵ cells per well). The transfections were performed using Lipofectamine 2000 (Life Technologies) according to the manufacturer’s protocol with CRISPR (2 μg); pPNT-HPRT (2 μg), CRISPR and pPNT or pPNT-HPRT constructs (2 μg + 2 μg). The medium was changed after 6 h.
The neomycin selection (350 μg/ml) (Sigma) was added 24/48 h after transfection and the resistant cells were pooled and analyzed by FISH.
The E14 neomycin resistant cells were seeded at a low density (1 × 10⁴ cells/well) and subjected to neomycin or neomycin and 6-TG media (30 μM) (Sigma Aldrich). After 3–4 days the resistant colonies were counted.

Paulis M., Castelli A., Lizier M., Susani L., Lucchini F., Villa A, & Vezzoni P. (2015). A pre-screening FISH-based method to detect CRISPR/Cas9 off-targets in mouse embryonic stem cells. Scientific Reports, 5, 12327.

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Maternal Antibiotics Influence Offspring Diabetes

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We treated pregnant (plugged) NOD mice with neomycin or vancomycin (all from Sigma) via drinking water at a final concentration of 1mg/ml for neomycin, 0.5mg/ml for vancomycin. The treatment was withdrawn within 24 hours after birth and diabetes development was observed in the offspring. The offspring from neomycin- or vancomycin-treated mothers were designated as Neo- or Van-mice.

Hu Y., Jin P., Peng J., Zhang X., Wong F.S, & Wen L. (2016). Different immunological responses to early-life antibiotic exposure affecting autoimmune diabetes development in NOD mice. Journal of autoimmunity, 72, 47-56.

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Auditory Tissue Culture and Viral Transduction

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The cochleae from WT P1 ICR mice were dissected, and the organ of Corti was cultured in DMEM/F12 (ThermoFisher Scientific) with 1% N2 supplement (ThermoFisher Scientific), 2% B27 supplement (ThermoFisher Scientific), and 50 mg/mL ampicillin (Sigma-Aldrich, St. Louis, MO) at 37°C with 5% CO₂. For the viral transduction, 2 h after cochlear explants, AAV-CasRx-g3 (3 × 10¹² vg/mL) was co-cultured with cochlear for 24 h. The medium was then changed to fresh DMEM/F12 mixture with 1 mM neomycin (Sigma-Aldrich), and cultured for 6 h. After 24 h of recovery in DMEM/F12 medium without neomycin, the cochlear explants were harvested for immunohistochemical staining.

Guo Y., Han L., Han S., Tang H., Wang S., Cui C., Chen B., Li H, & Shu Y. (2022). Specific knockdown of Htra2 by CRISPR-CasRx prevents acquired sensorineural hearing loss in mice. Molecular Therapy. Nucleic Acids, 28, 643-655.

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Antibiotic-induced Gut Microbiome Modulation

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A group of wild-type C3H/HeOuJ background mice were treated with antibiotic containing water ad libitum or control group was given standard drinking water. Antibiotic treatment consisted of neomycin and polymyxin B (final concentrations: 0.6 mg/mL for neomycin and 120 units/mL for polymyxin B, both from Sigma-Aldrich, St. Louis, MO, United States) given in drinking water for 8 weeks as described earlier (Prakash et al., 2015 (link)). This combination of antibiotics was chosen specifically for the ability of both to remain within the gut and not be absorbed into circulation; by using this combination to target the gut microbiome, we could focus primarily on its effects without disruption of other microbiomal niches. Supplementary Figure S6 demonstrates the effects of this combination on the richness, evenness, and diversity of the microbiome after 7 weeks of treatment.

Tian X., Hellman J., Horswill A.R., Crosby H.A., Francis K.P, & Prakash A. (2019). Elevated Gut Microbiome-Derived Propionate Levels Are Associated With Reduced Sterile Lung Inflammation and Bacterial Immunity in Mice. Frontiers in Microbiology, 10, 159.

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Titration and Detection of ASFV in Insect Cells

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The titers of infectious ASFV in serum and clarified spleen suspensions kept in the insect incubator at 27 °C and in the Thermomixer in the laboratory 27 °C were determined by end-point titration in PPAM as described above.
The presence of infectious virus in six selected T. molitor homogenate samples from study T2 containing ASFV DNA (with Cq values from 29.8 to 32.8) was determined using PPAM. The cells were maintained in MEM with 5% fetal bovine serum (FBS) and seeded in 24 well NUNC 24-well plates (Thermo Fisher Scientific). Larval homogenate (100 µL) was mixed with 100 µL MEM with 10% FBS, streptomycin (Sigma-Aldrich), neomycin (Sigma-Aldrich), amphotericin (Sigma-Aldrich) and benzylpenicillin (Sigma-Aldrich) and inoculated onto 1 mL cells (1,600,000 cells/mL). The inoculum was removed after one hour incubation at 37 °C (in 5% CO₂) and the cells were washed twice using 1x PBS. Following washing, 1 mL MEM containing 5% FBS, streptomycin, neomycin, amphotericin (Sigma-Aldrich) and benzylpenicillin was added to the cells.
After three days of incubation at 37 °C (in 5% CO₂), the cells were harvested by freezing and 100 µL of the 1st passage material was inoculated onto 1 mL fresh PPAM. After three days of incubation, virus-infected cells were identified using the IPMA as described above (Section 2.1.1).

Olesen A.S., Lazov C.M., Lecocq A., Accensi F., Jensen A.B., Lohse L., Rasmussen T.B., Belsham G.J, & Bøtner A. (2022). Uptake and Survival of African Swine Fever Virus in Mealworm (Tenebrio molitor) and Black Soldier Fly (Hermetia illucens) Larvae. Pathogens, 12(1), 47.

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