Neomycin

Manufactured by GoldBio

Sourced in United States

Neomycin is a broad-spectrum aminoglycoside antibiotic used in various laboratory applications. It functions as an inhibitor of protein synthesis, effectively suppressing the growth of a wide range of bacterial species.

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4 protocols using neomycin

Antibiotic Cocktail Modulates Gut Microbiome

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We chose an ABX cocktail with limited bioavailability that had previously been shown to broadly reduce gut microbiota populations in adult rodents without entering circulation and directly affecting host tissues when administered orally (Bercik et al., 2011 (link); Kiraly et al., 2016 (link)). The ABX cocktail, consisting of 0.5 mg/ml bacitracin (Sigma, USA), 2.0 mg/ml neomycin (GoldBio, USA), and 0.2 mg/ml vancomycin (Thermo Fisher, USA) was added to drinking water for the duration of the experiment. The anti-fungal pimaricin (1.2 μg/ml) was also added to the ABX cocktail solution, in order to decrease gut fungal overgrowth due to prolonged antibiotic use. SB (Sigma, USA) was provided to the mice ad libitum. SB solution was prepared in drinking water at concentration of 8 mg/ml. For ABX and SB (ABX+SB) co-treatment, SB was added to freshly prepared ABX solution. The SB and ABX+SB solutions were pH matched to ABX solution in order to control for acidity effects on consumption. All solutions were freshly prepared every 2 days.

Reyes R.E., Gao L., Zhang Z., Davies D.L, & Asatryan L. (2022). SUPPLEMENTATION WITH SODIUM BUTYRATE PROTECTS AGAINST ANTIBIOTIC-INDUCED INCREASES IN ETHANOL CONSUMPTION BEHAVIOR IN MICE. Alcohol (Fayetteville, N.Y.), 100, 1-9.

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Fluorescent Imaging Probe Preparation

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FDGlcU was purchased from Thermo Fisher Scientific (Waltham, MA, U.S.A.) and reconstituted in DDW. The eβG inhibitor was purchased from Merck (Cat# 347423) (Darmstadt, Germany), reconstituted in DMSO at 4.255 mg/mL, and then further diluted with DMSO for in vitro assay or diluted with DDW to 0.2 mg/mL for in vivo imaging. Isoflurane was purchased from Aesica (Queenborough, U.K.). Fluorescein was purchased from Sigma-Aldrich (St. Louis, MO, U.S.A.), reconstituted in DMSO, and then diluted with DDW before use. Ampicillin, neomycin, and vancomycin were purchased from Gold Biotechnology (St. Louis, MO, U.S.A.).

Chen M., Cheng K.W., Chen Y.J., Wang C.H., Cheng T.C., Chang K.C., Kao A.P, & Chuang K.H. (2017). Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe. Scientific Reports, 7, 3142.

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Multicolor Flow Cytometry of Immune Cells

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The antibiotics vancomycin, polymyxin B, ampicillin, and neomycin were obtained from Gold Biotechnology, Inc. (St. Louis, MO), and metronidazole was from Sigma-Aldrich Corp. (St. Louis, MO). Liposomal clodronate and control liposomes were obtained from Encapsula NanoSciences LLC (Brentwood, TN). The following reagents and anti-mouse antibodies were used for flow cytometry: Zombie Violet^TM fixable viability kit (BioLegend, San Diego, CA), CD45 (clone 30-F11, BioLegend), CD11b (clone M1/70, BioLegend), CD11c (clone N418, BioLegend), CD103 (clone 2E7, eBioscience/ Thermo Fisher Scientific, Waltham, MA), CD64 (clone X54-5/7.1, BioLegend), I-A/I-E (MHCII, clone M5/114.15.2, BioLegend), Ly6C (clone HK1.4, BioLegend), CD3e (clone 145–2 C11, BioLegend), CD4 (clone GK1.5, BioLegend), FoxP3 (clone FJK-16s, eBioscience), IFNγ (clone XMG1.2, BioLegend), IL17A (clone TC11-18H10.1, BioLegend), CD127 (clone A7R34, BioLegend), CD90.2 (Thy1.2, clone 30-H12, BioLegend), and RORγt (clone B2D, eBioscience).

Redhu N.S., Bakthavatchalu V., Conaway E.A., Shouval D.S., Tsou A., Goettel J.A., Biswas A., Wang C., Field M., Muller W., Bleich A., Li N., Gerber G.K., Bry L., Fox J.G., Snapper S.B, & Horwitz B.H. (2017). Macrophage dysfunction initiates colitis during weaning of infant mice lacking the interleukin-10 receptor. eLife, 6, e27652.

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Broad-spectrum antibiotic modulation of gut microbiota

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We chose a broad-spectrum antibiotic cocktail treatment (ABX) that leads to efficient depletion and dramatic alterations in the composition of the gut microbiota in adult rodents without entering circulation and directly affecting host tissues due to its lower absorbance [57 (link),58 (link)]. The ABX cocktail consisted of 0.5 mg/mL bacitracin (Sigma, St. Louis, MO, USA), 2.0 mg/mL neomycin (GoldBio, St. Louis, MO, USA), and 0.2 mg/mL vancomycin (Thermo Fisher, Waltham, MA, USA). It was provided to mice ad libitum in the drinking water throughout the duration of the study. Anti-fungal pimaricin (1.2 ug/mL, Molekula Group, Irvine, CA, USA) was added to the ABX cocktail, in order to decrease gut fungal overgrowth due to prolonged antibiotic use. SB (Sigma, St. Louis, MO, USA) was provided ad libitum in the drinking water with/without ABX. SB solution of 8 mg/mL in drinking water was freshly prepared every 2 days. SB and ABX+SB solutions were pH matched with the ABX solution to control for acidity effects on daily consumption.

Gao L., Davies D.L, & Asatryan L. (2022). Sodium Butyrate Supplementation Modulates Neuroinflammatory Response Aggravated by Antibiotic Treatment in a Mouse Model of Binge-like Ethanol Drinking. International Journal of Molecular Sciences, 23(24), 15688.

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