Antibiotic antimycotic solution

Manufactured by Lonza

Sourced in United States, Switzerland

Antibiotic/antimycotic solution is a sterile, liquid mixture of antibiotics and antifungal agents commonly used in cell culture applications to prevent bacterial and fungal contamination. The solution contains a combination of antibiotics and antimycotics, which inhibit the growth of a broad spectrum of microorganisms.

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Antibiotic/antimycotic (32 citations) Antibiotics/Antimycotics (2 citations) Antibiotic solution (2 citations)

14 protocols using antibiotic antimycotic solution

Cell Culture of TCam-2 and HEK293FT

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TCam-2 cells were cultured in RPMI with GlutaMAX medium (Gibco #61870044) supplemented with 10% (v/v) fetal bovine serum (HyClone #sv 30160.03) and 1% (v/v) antibiotic/antimycotic solution (Lonza #17-602E). HEK293FT cells were cultured in low glucose DMEM medium (Lonza #BE12-707F) supplemented with 2 mM L-Glutamine (Thermo Fisher Scientific, Waltham, MA, USA, #25030081), 10% (v/v) fetal bovine serum (HyClone), and 1% (v/v) antibiotic antimycotic solution (Lonza).

Ilaslan E., Kwiatkowska K., Smialek M.J., Sajek M.P., Lemanska Z., Alla M., Janecki D.M., Jaruzelska J, & Kusz-Zamelczyk K. (2022). Distinct Roles of NANOS1 and NANOS3 in the Cell Cycle and NANOS3-PUM1-FOXM1 Axis to Control G2/M Phase in a Human Primordial Germ Cell Model. International Journal of Molecular Sciences, 23(12), 6592.

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Culturing and Transfecting TCam-2 Cells

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TCam-2 cells (supplied by our collaborator Dr Kitazawa) were cultured in RPMI with GlutaMAX medium (Life Technologies, Poland) supplemented with 10% (v/v) fetal bovine serum (FBS; HyClone, USA) and 1% (v/v) antibiotic/antimycotic solution (Lonza, Switzerland). Cells were transfected with plasmids or siRNA using the Neon Transfection System (Life Technologies), according to the manufacturer’s protocol, followed by culture in the same medium without antibiotic/antimycotic solution.

Janecki D.M., Sajek M., Smialek M.J., Kotecki M., Ginter-Matuszewska B., Kuczynska B., Spik A., Kolanowski T., Kitazawa R., Kurpisz M, & Jaruzelska J. (2018). SPIN1 is a proto-oncogene and SPIN3 is a tumor suppressor in human seminoma. Oncotarget, 9(65), 32466-32477.

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Immortalized Cell Lines for ATP Synthase and ANT Studies

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Immortalized HAP 1 and MEF cell lines were used for this study.⁶ (link)^,⁸ (link) HAP 1 cells were grown Iscove’s Modified Dulbecco’s Medium (IMDM), supplemented with 10% Heat-Inactivated Fetal Bovine Serum (HI FBS; Life Technologies), 10 mL per L of Antibiotic Antimycotic Solution (Penicillin/Streptomycin/Amphoterichin B; Sigma Aldrich) and 2 mM L-Glutamine. MEF cells were grown in high-glucose Dulbecco’s Modified Eagle Medium (DMEM; Cytiva) supplemented with 10% HI FBS, 10 mL per L of Antibiotic Antimycotic Solution, and 1X Non-Essential Amino acids (NEAA; Lonza). HAP 1 Δ (c+δ) cell line that lacks c and δ-subunits of ATP synthase was used for the study of the role of ATP synthase in high-conductance PTP. MEF ANT Triple KO cell line lacking 3 ANT genes was used to study the role of ANT in high conductance PTP. MEF ANT Triple KO cells were grown in the same media as MEF WT cells with the addition of 1mM Sodium Pyruvate (Gibco) and 25mg/500mL Uridine (Sigma Aldrich). Cells were maintained in a humidified cell incubator, at 37°C under a 5% CO2 atmosphere.

Neginskaya M.A., Morris S.E, & Pavlov E.V. (2022). Both ANT and ATPase are essential for mitochondrial permeability transition but not depolarization. iScience, 25(11), 105447.

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Murine Stromal Cell Culture Protocol

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All cultures with stromal cells were carried out with murine stromal cells harvested from femurs and tibias and cultured through one passage (P1) and supplemented with Gibco MEMα, nucleosides, No Phenol Red Media supplemented with 100 U/mL penicillin-streptocycin, 100 U/mL antibiotic antimycotic solution (Lonza BioWhittaker Antibiotics: Penicillin-Streptomycin Mixtures) and 20% FBS (Denville). Media and cytokines were replaced every 3 days.

Camacho V., Matkins V.R., Patel S.B., Lever J.M., Yang Z., Ying L., Landuyt A.E., Dean E.C., George J.F., Yang H., Ferrell P.B., Maynard C.L., Weaver C.T., Turnquist H.R, & Welner R.S. (2020). Bone marrow Tregs mediate stromal cell function and support hematopoiesis via IL-10. JCI Insight, 5(22), e135681.

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In Vitro T. gondii Tachyzoite Stimulation Assay

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Heparinized blood samples were processed within 2 h of collection by mixing 500 μL blood with 500 μL RPMI 1640 medium (Gibco, Paisley, UK) supplemented with 10% foetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, USA) and 1% antibiotic/antimycotic solution (Lonza, Basel, Switzerland). Blood cells were cultured in 24-well plates (Thermo Fisher Scientific, Waltham, USA) in the presence of 10⁶ lyophilized T. gondii tachyzoites (see Parasites and inoculum) as antigen, concanavalin A as a control (ConA, Sigma−Aldrich, Madrid, Spain), both at final concentrations of 5 μg/mL, or PBS as a negative control. The plates were incubated in a 5% CO₂/37°C/100% humidity atmosphere for 48 h and centrifuged at 1000×g for 10 min at 4°C. Finally, cell-free culture supernatants were stored at −80°C until analysis.

Largo-de la Torre A., Diezma-Díaz C., Calero-Bernal R., Atencia-Cibreiro G., Sánchez-Sánchez R., Ferre I., Regidor-Cerrillo J, & Ortega-Mora L.M. (2022). Archetypal type II and III Toxoplasma gondii oocysts induce different immune responses and clinical outcomes in experimentally infected piglets. Frontiers in Immunology, 13, 1021556.

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Porcine Aortic Valve Cell Isolation

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Primary VICs were harvested from aortic valve leaflets dissected out of fresh porcine hearts from a commercial abattoir (Fisher Ham and Meats, Spring, TX, USA). The leaflets were incubated with collagenase type II (550 U/ml, 30 min; Worthington Biochemical, Lakewood, NJ, USA), followed by manually scraping off endothelial cells and mincing the leaflets, which were then digested with collagenase type III (260 U/ml, 4 h; Worthington) in an incubated shaker (Stephens et al., 2007 (link)). Isolations from six different donors were pooled together to compensate for biological variability. The cells were cultured on tissue culture polystyrene flasks in growth medium [GM; 1:1 Dulbecco’s modified Eagle’s medium (DMEM; Corning, Tewksbury, MA, USA):F12 (Hyclone, Thermo Fisher Scientific, Waltham, MA, USA) with 10% bovine growth serum (BGS; Hyclone), 1.6% 1-m-4-(2-hydroxyethyl)-1- piperazine-ethanesulphonic acid (HEPES; Thermo Fisher Scientific) and 1% antibiotic–antimycotic solution (Lonza, Walkerville, MD, USA)] before being encapsulated. After encapsulation, cell-laden hydrogels were cultured in GM for 1 day and changed to different medium conditions, including GM, or GM supplemented with 50 μg/ml AA (GM + AA), or osteogenic medium (Osteo M; GM supplemented with 50 μg/ml AA, 10mM β-glycerophosphate disodium and 10 nM dexamethasone).

Wu Y., Puperi D.S., Grande-Allen K.J, & West J.L. (2015). Ascorbic acid promotes extracellular matrix deposition while preserving valve interstitial cell quiescence within 3D hydrogel scaffolds. Journal of tissue engineering and regenerative medicine, 11(7), 1963-1973.

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Isolation and Culture of Porcine Aortic Valve Interstitial Cells

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As previously described ^{50 (link)}, primary VICs were harvested from aortic valve leaflets
dissected out of fresh porcine hearts from a commercial abattoir (Fisher Ham and
Meats, Spring, TX, USA). Briefly, the leaflets were incubated with collagenase
type II (550 U/ml, 30 min; Worthington Biochemical Corp, Lakewood, NJ) followed
by manually scraping off endothelial cells and mincing the leaflets, which were
then digested with collagenase type III (260 U/ml, 4 h; Worthington) in an
incubated shaker. Isolations from six different donors were pooled together to
compensate for biological variability. Cells were cultured on tissue culture
polystyrene flasks in VIC growth media [1:1 DMEM (Corning, Tewksbury, MA,
USA):F12 (Hyclone, Thermo Fisher Scientific, Waltham, MA, USA) with 10% bovine
growth serum (BGS; Hyclone), 1.6% 1 M
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES; Thermo Fisher
Scientific), and 1% antibiotic-antimycotic solution (Lonza, Walkerville, MD)]
before being seeded. VICs between passage 2 and 4 were used for study.

Wu Y., Grande-Allen K.J, & West J.L. (2016). Adhesive Peptide Sequences Regulate Valve Interstitial Cell Adhesion, Phenotype and Extracellular Matrix Deposition. Cellular and molecular bioengineering, 9(4), 479-495.

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Wound Healing Assay with HaCaT and DOK Cells

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HaCaT and DOK cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium)/Ham F12 (Sigma-Aldrich Chemie Gmbh, Munich, Germany) and supplemented with 10% fetal bovine serum (FBS), and antibiotic-antimycotic solution (both from Lonza, Basel, Switzerland). Cells were seeded in 35 mm glass bottom petri dishes (7.5 × 10⁵ cells/dish), for time-lapse videomicroscopy wound healing experiments, or in 16 wells E-plates (7.5 × 10³ HaCaT cells/well, and 15 × 10³ DOK cells/well, respectively) for time-lapse impedance monitoring. To prepare culture surfaces for the experiments, three different matrix proteins were used: collagen type I, from calf skin (C9791, Sigma-Aldrich Chemie Gmbh); fibronectin from bovine plasma (33010-018, Invitrogen Molecular Probes, Eugene, OR, USA); and laminin from mouse basement membrane (354232, BD Bioscience, San Jose, CA, USA).

Niculiţe C.M., Nechifor M.T., Urs A.O., Olariu L., Ceafalan L.C, & Leabu M. (2018). Keratinocyte Motility Is Affected by UVA Radiation—A Comparison between Normal and Dysplastic Cells. International Journal of Molecular Sciences, 19(6), 1700.

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Avian Reovirus Isolation from Tissue

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Each organ sample was homogenized individually in sterile phosphate buffered saline using Qiagen TissueLyser II at 50 Hz for 3–5 min. After centrifugation at 10,000× g (for 5 min at 4 °C) supernatants were filtered through a 0.45 µm PES syringe filter. For virus isolation, filtered homogenates were inoculated in a four-member ten-fold dilution on chicken hepatocellular carcinoma cell line LMH (Leghorn male hepatoma, ATCC CRL-2117). LMH cells were cultured in DMEM medium (Lonza) supplemented with 5% fetal bovine serum (Gibco) and 1% antibiotic/antimycotic solution (Lonza) in a 37 °C incubator with constant supply of 5% CO₂. Upon giant cell formation, the cytopathogenic effect (CPE) characteristic of ARV, could be observed (typically after 5–6 days), the isolates were propagated in a 25 cm² cell culture flask. When CPE was not seen following the incubation period, blind passages were carried out; after three blind passages, CPE-negative samples were discarded.

Gál B., Varga-Kugler R., Ihász K., Kaszab E., Domán M., Farkas S, & Bányai K. (2023). Marked Genotype Diversity among Reoviruses Isolated from Chicken in Selected East-Central European Countries. Animals : an Open Access Journal from MDPI, 13(13), 2137.

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Synthesis and Characterization of Magnetic Nanoparticles

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To synthesize the nanoparticles, analytical grade iron chloride hexahydrate (FeCl₃·6H₂O), manganese chloride tetrahydrate (MnCl₂·4H₂O) from Sigma Aldrich were used as initial precursors. Ammonium hydroxide solution (NH₄OH solution; 25%) from Sigma Aldrich was used as the reducing agent.
For cellular studies, normal murine fibroblasts cell line (L929) and human glioma cell line (U87-MG) were purchased from NCCS, Pune, India. The cells were cultured in Dulbecco's Modified Eagles Medium (Hyclone) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco) and 1% antibiotic–antimycotic solution (Lonza). The cell cultures were grown at 37 °C under 5% CO₂.

Gupta R., Tomar R., Chakraverty S, & Sharma D. (2021). Effect of manganese doping on the hyperthermic profile of ferrite nanoparticles using response surface methodology. RSC Advances, 11(28), 16942-16954.

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