Gentamycin sulfate

Manufactured by Sartorius

Sourced in Israel

Gentamycin sulfate is a laboratory product used as a selective antibiotic agent in cell culture media. It is an aminoglycoside antibiotic derived from the bacterium Micromonospora purpurea, and is effective against a wide range of Gram-negative and some Gram-positive bacteria.

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Lab products found in correlation

Fetal bovine serum (fbs), by Sartorius (5 mentions) Sodium pyruvate, by Sartorius (4 mentions) Low glucose dmem, by Sartorius (4 mentions) L glutamine, by Merck Group (3 mentions) L glutamine, by Sartorius (3 mentions) Dulbecco modified eagle medium (dmem), by Sartorius (2 mentions) Amphotericin b, by Sartorius (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Amphotericin, by Sartorius (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Dulbecco s modified eagle medium dmem, by Sartorius (2 mentions) Dmem f12, by Sartorius (2 mentions) Pdgf aa, by R&D Systems (1 mentions) Glutamine, by Merck Group (1 mentions) Fibroblast growth factor 2 (fgf2), by R&D Systems (1 mentions) Hoescht 33342, by Merck Group (1 mentions) Lsm 700, by Zeiss (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions) Crl 3216, by American Type Culture Collection (1 mentions)

Spelling variants of this product

Gentamycin (17 citations) Gentamycin Sulfate Solution (2 citations)

13 protocols using gentamycin sulfate

Isolation and Culture of Oligodendrocyte Precursor Cells

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Primary OPC cultures were isolated from naïve P0 to P1 neonatal C57/BL6 mice cortices as previously described by Chen et al. (2007 (link)), with minor modifications (Barateiro & Fernandes, 2014 (link)). Briefly, a mixed glial culture isolated from neonatal mice was grown for 8 days in Dulbecco's modified Eagle's medium (DMEM) low glucose (Biological Industries, Israel) supplemented with 5% fetal bovine serum (Biological Industries, Israel), 1 mM sodium pyruvate (Biological Industries, Israel), 1 mM L‐glutamine (Sigma‐Aldrich, Israel) and 0.6% Gentamycin Sulfate (Biological Industries, Israel). Culture medium was replaced every 2 days. After 8 days, microglia were detached by 30 min shaking at 140 rpm using an orbital shaker. After medium was removal, a new fresh culture medium was added, and OPCs at the top of the astrocyte monolayer were detached by shaking for 18 h at 200 rpm. Cells were plated in DMEM/F‐12 (Rhenium, Israel) supplemented with 1% B27 supplement (Rhenium, Israel) and 0.6% Gentamycin Sulfate (Biological Industries, Israel). Fibroblast growth factor‐2 (FGF‐2; 20 ng/ml; R&D systems) and Platelet‐derived growth factor (PDGF‐AA; 20 ng/ml; R&D systems) were added to cultures daily unless otherwise specified. Timeline is depicted in Figure S1a.

Zveik O., Fainstein N., Rechtman A., Haham N., Ganz T., Lavon I., Brill L, & Vaknin‐Dembinsky A. (2022). Cerebrospinal fluid of progressive multiple sclerosis patients reduces differentiation and immune functions of oligodendrocyte progenitor cells. Glia, 70(6), 1191-1209.

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Imaging 4T1 cells infected with Salmonella

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1×10⁵ cells/well (2 mL) of triple negative breast cancer mouse cell-line (4T1; ATCC CRL2539) were seeded in 6-well plates in a RPMI 1640 medium (Sigma-Aldrich), supplemented with 10% heat inactivated Fetal Bovine Serum (Biological Industries, Beit-Haemek, Israel), 2 mM of L-Glutamine Solution, 10 U/mL of penicillin G sodium salt and 0.01 mg/mL of streptomycin sulfate for 24 hours at 37 °C in a 5% CO₂ humid atmosphere. The following day, the 4T1 cells were washed, to remove remnants of antibiotics, and Salmonella at multiplicity of infection (MOI) 100:1 was added for 3h. The 4T1 cells were washed with PBS and gentamycin sulfate at concentration of 20μg/mL (Biological industries, Israel) to remove external bacteria. Cells were incubated with Hoescht 33342 (Sigma-Aldrich) and Dil (life technologies, Israel) to stain the nucleus and the cell’s membrane, respectively. Images were obtained using Confocal Laser Scanning Microscopy Zeiss LSM 700 and ZEN imaging software integrated to a three dimensional (Z-stack) image.

Zoaby N., Shainsky-Roitman J., Badarneh S., Abumanhal H., Leshansky A., Yaron S, & Schroeder A. (2016). Autonomous bacterial nanoswimmers target cancer. Journal of controlled release : official journal of the Controlled Release Society, 257, 68-75.

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Cytotoxicity Evaluation of Cultured Fibroblasts

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Cytotoxicity of solutions was evaluated on cultured hPDL fibroblast cells in Research Laboratory, Selcuk University, Faculty of Dentistry, Konya, Turkey. The cells were grown in 96-well polystyrene plates containing Dulbecco's modified Eagle's medium (DMEM) (Biological Industries, Kibbutz Beik Haemek, Israel), supplemented with 10% fetal bovine serum (FBS) (Biological Industries, Kibbutz Beik Haemek, Israel), 250 μg/ml gentamycin sulfate, (Biological Industries, Kibbutz Beik Haemek, Israel), 5 μg/ml amphotericin B (Biological Industries, Kibbutz Beik Haemek, Israel), and were incubeted in a humidified atmosphere, 95% air and 5% CO₂ at 37°C for 24 h in water based incubator (NUAIRE, Fernbrook Lane N Plymouth, USA).

Ok E., Adanir N, & Hakki S. (2015). Comparison of cytotoxicity of various concentrations origanum extract solution with 2% chlorhexidine gluconate and 5.25% sodium hypochlorite. European Journal of Dentistry, 9(1), 6-10.

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Isolation of Mouse Primary Astrocytes

Cited 1 time

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Mouse primary astrocytes were isolated from naïve P0 to P1 neonatal C57/BL6 mice cortices as previously described by Chen et al. [84 (link)], with minor modifications [85 (link),86 (link)]. Briefly, a mixed glial culture isolated from neonatal mice was grown for 8 days in Dulbecco’s modified Eagle’s medium (DMEM) low glucose (Biological Industries, Kibbutz Beit-Haemek, Israel) supplemented with 5% fetal bovine serum (Biological Industries, Kibbutz Beit-Haemek, Israel), 1 mM sodium pyruvate (Biological Industries, Kibbutz Beit-Haemek, Israel), 1 mM L-glutamine (Sigma–Aldrich, Rehovot, Israel) and 0.6% Gentamycin Sulfate (Biological Industries, Kibbutz Beit-Haemek, Israel). Culture medium was replaced every 2 days. After 8 days, microglia were detached by 30 min shaking at 140 rpm using an orbital shaker. After medium was removal, a new fresh culture medium was added, and OPCs at the top of the astrocyte monolayer were detached by shaking for 18 h at 200 rpm. Media was replaced and astrocytes were grown for further 7 days. Cells were detached from flasks using TrypLE (Thermo Fisher Scientific, Waltham, MA, USA). All cultures expressed high level of GFAP (mean of 96.6% GFAP+ cells, Figure S1).

Zveik O., Rechtman A., Haham N., Adini I., Canello T., Lavon I., Brill L, & Vaknin-Dembinsky A. (2022). Sera of Neuromyelitis Optica Patients Increase BID-Mediated Apoptosis in Astrocytes. International Journal of Molecular Sciences, 23(13), 7117.

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Culturing Mouse and Human Cell Lines

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NIH3T3 (Mouse embryo fibroblast), RAW (RAW267.4, Murine monocytes/macrophages-like) and 293FT (Human embryonal kidney) were obtained from ATCC, Manassas, Virginia, USA (CRL-1658, TIB-71 and CRL-3216, respectively). These cell lines were maintained in DMEM supplemented with 10% FCS, 2.5 μg/ml Amphotericin and 50 μg/ml Gentamycin Sulfate (Biological Industries, Beit-Haemek, Israel). Mouse iPS cell line (miPS-B6-GFP) was provided by Prof. Lior Gepstein. Undifferentiated colonies were cultured on mitotically inactivated mouse embryonic fibroblasts (MEF) feeder layer, as previously described [11 (link)]. Cells were maintained in DMEM supplemented with 15% FCS (Biological Industries), 0.1% leukemia inhibitory factor (LIF) (Millipore), 1mM L-glutamine, 0.1mM Mercaptoethanol, and 1% nonessential amino acid stock (all from Invitrogen).

Khateb M., Fourier N., Barnea-Yizhar O., Ram S., Kovalev E., Azriel A., Rand U., Nakayama M., Hauser H., Gepstein L, & Levi B.Z. (2016). The Third Intron of the Interferon Regulatory Factor-8 Is an Initiator of Repressed Chromatin Restricting Its Expression in Non-Immune Cells. PLoS ONE, 11(6), e0156812.

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PBMC Isolation Using Density Gradient

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PBMCs were isolated by density gradient separation using LymphoPrep (cat #1114547; Axis-Shield, Oslo, Norway) at 1200 x g for 30 minutes, washed twice in phosphate buffered saline followed by centrifugation at 400 x g, and re-suspended in RPMI 1640 (cat #01-106-1a; Biological Industries, Kibbutz Beit Haemek, Israel) containing 30% (v/v) AB serum, 25 mM HEPES, 2 mM L-glutamine (cat #25030–024; Gibco, Life Technologies, Carlsbad, CA), and 50 μg/mL gentamycin sulfate (cat #03-035-1c; Biological Industries). All subsequent centrifugations were carried out at 400 x g. Human serum isolated from healthy male donors of blood group AB (AB serum) was purchased from Lonza (cat # 14-490E; Basel, Switzerland) and used as the serum source in all experiments.

Kristensen B., Hegedüs L., Lundy S.K., Brimnes M.K., Smith T.J, & Nielsen C.H. (2015). Characterization of Regulatory B Cells in Graves’ Disease and Hashimoto’s Thyroiditis. PLoS ONE, 10(5), e0127949.

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Neuroinflammation Inhibition Protocol

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Ladostigil was a gift from Spero Biopharma (Jerusalem, Israel). Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12) was used. Gentamycin sulfate and L-Glutamine were obtained from Biological Industries (Beit-Haemek, Israel) and BzATP, bovine serum albumin (BSA), and LPS were from Escherichia coli 055:B5, purified by trichloracetic acid extraction from Sigma-Aldrich Israel Ltd. (Rechovot, Israel).

Reichert F., Zohar K., Lezmi E., Eliyahu T., Rotshenker S., Linial M, & Weinstock M. (2024). Ladostigil Reduces the Adenoside Triphosphate/Lipopolysaccharide-Induced Secretion of Pro-Inflammatory Cytokines from Microglia and Modulate-Immune Regulators, TNFAIP3, and EGR1. Biomolecules, 14(1), 112.

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Astrocytes Exposed to NMO Sera

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Cells were seeded in plates for 24 h. Then, media was replaced for DMEM low glucose (Biological Industries, Kibbutz Beit-Haemekm Israel) supplemented with 1 mM sodium pyruvate (Biological Industries, Kibbutz Beit-Haemek, Israel), 1 mM L-glutamine (Sigma–Aldrich, Rehovot, Israel) and 0.6% Gentamycin Sulfate (Biological Industries, Kibbutz Beit-Haemek, Israel). Human sera of either NMO patients or HCs (20% of media) were added into mouse primary astrocytes media for 48 h. Then, DNA damage response was evaluated using anti-H2AX (Cat# sc-517348, Santa Cruz biotechnology Inc., Dallas, TX, USA, 1:100). Each assay was repeated independently at least three times.

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Culturing Human Cancer Cell Lines

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Human cell lines including PANC-1 (pancreatic cancer), OV-90 (ovarian cancer) and MDA-MB-231 (breast adenocarcinoma) were obtained from American Type Culture Collection and used within 6 months. Cell lines were maintained in DMEM (Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FBS; Biological Industries, Beit Haemek, Israel), 50 µg/ml gentamycin sulfate (Biological Industries) and 250 µg/ml amphotericin B (Biological Industries).

Ohana J., Sandler U., Kass G., Stemmer S.M, & Devary Y. (2017). dTCApFs, a derivative of a novel human hormone peptide, induces apoptosis in cancer cells through a mechanism involving loss of Golgi function. Molecular and Clinical Oncology, 7(6), 991-999.

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Culturing Mouse and Human Cell Lines

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NIH3T3 (mouse embryo fibroblast) and 293FT (human embryonal kidney) were obtained from ATCC (Manassas, VA, USA) (CRL-1658 and CRL-3216, respectively). These cell lines were maintained in DMEM supplemented with 10% fetal calf serum (FCS), amphotericin (2.5 µg/mL) and gentamycin sulfate (50 µg/mL, Biological Industries, Beit-Haemek, Israel).

Katsman M., Azriel A., Horev G., Reizel Y, & Levi B.Z. (2022). N-VEGF, the Autoregulatory Arm of VEGF-A. Cells, 11(8), 1289.

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