Antibiotics antimycotics

Manufactured by Merck Group

Sourced in United States, Germany

Antibiotics/antimycotics are laboratory products used to inhibit the growth of bacteria and fungi. They are commonly used in cell culture applications to prevent microbial contamination. These products contain a combination of antibiotics and antifungal agents to provide broad-spectrum protection.

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Antibiotic/antimycotic (245 citations) Antibiotic/antimycotic mix (20 citations) Antibiotic-antimycotic mixture (18 citations) Antibiotic/antimycotic cocktail (15 citations) Antibiotic Antimycotic Solution 100x (11 citations) Antimycotics (7 citations) Antibiotic/ antimycotic solution (AA; (7 citations) Antibiotics and antimycotics (5 citations) Antibiotic Antimycotic Solution (Ab/AM) (4 citations) Antibiotic-Antimycotic 100× (4 citations)

28 protocols using antibiotics antimycotics

Culturing Human Breast Cancer Cell Lines

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The human breast cancer cell lines MDA-MB231, MDA-MB438, BT474 and MCF7 cell lines were provided by American Type Culture Collection (ATCC, Manassas, VA, USA).
MDA-MB438 and MDA-MB231 were maintained in in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, USA) and 1% antibiotics/antimycotics (Life Technologies, Gaithersburg, MD, USA).
BT474 were cultured in Roswell Park Memorial Institute (RPMI, Sigma-Aldrich, Saint Louis, Missouri, USA) medium added with 10% FBS and 1% antibiotics/antimycotics.
MCF7 were grown in the base medium ATCC-formulated Eagle's Minimum Essential Medium, (ATCC, Manassas, VA, USA - Catalog No. 30-2003), complemented by 01 mg/ml human recombinant insulin (Sigma-Aldrich, Saint Louis, Missouri, USA), 10% FBS and 1% antibiotics/antimycotics.
KPL4 cell line was kindly provided by Dr Roberto Bianco at Federico II University (Napoli, Italy) and cultivated in DMEM medium with 5% FBS and 1% antibiotics/antimycotics.
The SUM159 cell lines were purchased from Asterand Inc. (Detroit, MI, USA) and were grown in Ham's F-12 medium supplemented with 5% heat-inactivated fetal bovine serum (FBS) and 1% antibiotics/antimycotics (Invitrogen, Carlsbad, CA, USA).
All cell lines were maintained in a humidified incubator with 5% CO2 atmosphere at 37^°C.

Morgillo F., Della Corte C.M., Diana A., Mauro C.D., Ciaramella V., Barra G., Belli V., Franzese E., Bianco R., Maiello E., de Vita F., Ciardiello F, & Orditura M. (2017). Phosphatidylinositol 3-kinase (PI3Kα)/AKT axis blockade with taselisib or ipatasertib enhances the efficacy of anti-microtubule drugs in human breast cancer cells. Oncotarget, 8(44), 76479-76491.

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C2C12 Mouse Myoblast Differentiation

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The C2C12 mouse myoblasts (ATCC: American Type Culture Collection, Manassas, VA) were cultured in proliferation medium [PM: Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco); 10% foetal calf serum (Gibco); 1% antibiotics/antimycotics (Sigma)]. Cells were put in plate of 8.8 cm² at a density of 2 × 10⁵ cells/ml in PM. When reaching 80–90% confluence, PM was changed toward differentiation medium [DM: DMEM (Gibco); 2% heat inactivated horse serum (Gibco); 1% antibiotics/antimycotics (Sigma)]. This medium change corresponds to day 0 of differentiation. Cells were cultured into an incubator at 37 °C, 5% CO₂ and saturated humidity, and culture media were changed every 2 days.

Claeyssen C., Bastide B, & Cieniewski-Bernard C. (2022). Global O-GlcNAcylation changes impact desmin phosphorylation and its partition toward cytoskeleton in C2C12 skeletal muscle cells differentiated into myotubes. Scientific Reports, 12, 9831.

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IGF1 Signaling in Lung Adenocarcinoma

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Human non-small cell lung adenocarcinoma cell lines A549 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotics/antimycotics (Sigma, St Louis, MO, USA). The 3 cell lines were routinely sub-cultured every 3–4 days. All siRNA transfections were performed using Dharmafect1 #T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per manufacturer’s protocol. After total 48 hours of transfection, cells were serum starved for 12 hours. After which IGF1 (50ng/ml) was added to stimulate the cells. For protein stability studies, Cycloheximide (20uM), and for testing degradation pathways, Monensin (10uM) were added an hour prior to stimulating with IGF1. At the end of stimulation for various time points, cells were harvested in CHAPS lysis buffer (1% CHAPS detergent, 150mM NaCl, 50mM Tris pH 7, 5mM EDTA) supplemented with protease and phosphatase inhibitors. Protein was quantified by using Pierce's BCA Protein Assay Reagent Kit (#23227) from Pierce Biotechnology, Rockford, IL, USA as per manufacturer’s protocol.

Kurlawala Z., Dunaway R., Shah P.P., Gosney J.A., Siskind L.J., Ceresa B.P, & Beverly L.J. (2017). Regulation of Insulin-like Growth Factor Receptors by Ubiquilin1. The Biochemical journal, 474(24), 4105-4118.

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Modulation of mouse brain endothelial cell SIRT3

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Mouse brain microvascular endothelial cells (mBMECs, from Angio-Proteomie) were cultured in DMEM/HG medium (ThermoFisher, Waltham, MA, USA) supplemented with 20% FBS (ThermoFisher, Waltham, MA, USA), and 1% Antibiotic–Antimycotic (100×) solution (ThermoFisher, Waltham, MA, USA). Cells were infected with lentivirus carrying three short hairpin RNA (shRNA) sequences against mouse Sirt3 or a scramble lentivirus and selected with puromycin for stable expression. Puromycin was added 3 days after infection, with a concentration of 1 μg/mL, which was replaced every 2–4 days depending on the confluency of the cells. Human umbilical vein endothelial cells (HUVECs, from Lonza, Visp, Switzerland) between passages 6–9 were cultured in DMEM/F12 medium supplemented with 10% FBS, 1% Antibiotics–Antimycotics, and endothelial cell growth supplement (Sigma-Aldrich, St. Louis, MO, USA). HUVECs were transfected with human SIRT3 siRNA (GenePharma, Suzhou, China) or universal scrambled negative control siRNA using Lipofectamine RNAiMAX Transfection Reagent (ThermoFisher, Waltham, MA, USA), according to the manufacturer’s instructions and were used within 36–48 h after transfection.

Cao X., Wu Y., Hong H, & Tian X.Y. (2022). Sirtuin 3 Dependent and Independent Effects of NAD+ to Suppress Vascular Inflammation and Improve Endothelial Function in Mice. Antioxidants, 11(4), 706.

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Evaluating Prdx6 Effects on Cell Lines with Varying TLR4 Levels

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Since Prdx6 is a TLR4 ligand [26 (link)], it can penetrate into the cell in the form of a complex (TLR4/Prdx6). Therefore, it was necessary to test exogenous Prdx6 (WT and S32A) in cell lines that differ in the level of TLR4 expression. For that purpose, we used 3T3 mouse embryonic fibroblast cells with an average level of TLR4 [39 (link)], RAW 264.7 mouse macrophage cells highly expressing TLR4 [40 (link)], and A549 human lung carcinoma cells which have nearly no TLR4 expression (which results in their low sensitivity to lipopolysaccharides) [34 (link)]. Cells were seeded in culture flasks (25 cm²) to a density of 1 × 10⁶ cells/flask in DMEM medium (Paneco, Moscow, Russia), supplemented with 10% fetal bovine serum–FBS (Thermo Fisher Scientific, Swindon, UK) and a mixture of antibiotics/antimycotics (Sigma- Aldrich, USA). Cell culturing was carried out at 37 °C with 5% CO₂. Passage 5–8 cells were used in the study. Cell cultures used in this work were regularly tested for mycoplasma infections (Figures S1–S3): using microscopic control, Hoechst 33342 staining (500 ng/mL) (Thermo Fisher Scientific, Eugene, OR, USA) [60 (link)] and using real-time PCR [61 (link)].

Sharapov M.G., Goncharov R.G., Parfenyuk S.B., Glushkova O.V, & Novoselov V.I. (2022). The Role of Phospholipase Activity of Peroxiredoxin 6 in Its Transmembrane Transport and Protective Properties. International Journal of Molecular Sciences, 23(23), 15265.

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In vitro Parasite Culture Protocol

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Isolated parasite blood stages were seeded into 96-well plates at a density of 200,000 cells per well and subsequently cultured in RPMI 1640 medium (Gibco, Life Technologies) at 26 °C with 5% CO₂. The medium was fortified with 20% heat-inactivated fetal bovine serum (Biosera Europe, France), 1 mL of GlutaMAX™ (ThemoFisher, USA), carp red blood cell lysate equalling 1:10 parasite cells: host cells (host cells lysed in water), and 0.008 mL of antibiotics/antimycotics (10,000 U/mL, Sigma-Aldrich, USA).

Kyslík J., Born-Torrijos A., Holzer A.S, & Kosakyan A. (2024). RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari. Scientific Reports, 14, 3545.

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Endothelial Cell Culture Protocol

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Human microvascular endothelial cells (HMECs) and Human umbilical vein endothelial cells (HUVECs) were cultured in fully supplemented EBM-2 endothelial growth media with 10% FBS and 1% Antibiotics – Antimycotics (Sigma, St. Louis, MO) and used at passages 7–12. Primary mouse endothelial cells (MECs) were cultured at 37°C, 5% CO2 in corresponding endothelial media with supplements including growth factors, 10% FBS and 1% Antibiotics – Antimycotics. HMEC cells were from Cell Systems (Kirkland, WA). HUVEC were from Lonza (Alpharetta, GA).

Mao M., Varadarajan S., Fukai T., Bakhshi F.R., Chernaya O., Dudley SC J.r., Minshall R.D, & Bonini M.G. (2014). Nitroglycerin Tolerance in Caveolin-1 Deficient Mice. PLoS ONE, 9(8), e104101.

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Bovine SMSC Isolation and Differentiation

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Bovine SMSCs were isolated form longissimus muscle of a 2-week-old Holstein bull calf, as previously described [16 (link)]. SMSCs were obtained by percoll digestion and differential centrifugation. SMSCs were expanded in growth medium consisting of Dulbecco’s Modified Eagle Medium (DMEM; Sigma, St. Louis, MO, USA), 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA), and 1% antibiotics-antimycotics (Sigma) at 37 °C under 5% CO₂. To induce differentiation, growth medium was replaced by differentiation medium consisting of DMEM, 2% horse serum (Hyclone, Logan, UT, USA), and antibiotics-antimycotics.

Zhu Y., Li P., Dan X., Kang X., Ma Y, & Shi Y. (2022). miR-377 Inhibits Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells by Targeting FHL2. Genes, 13(6), 947.

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Immunocytochemistry and Immunoblotting of Photoactive Cryptochromes

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N2A and HEK293 cells were maintained in DMEM with 10% fetal calf serum and 1% antibiotics/antimycotics (Sigma-Aldrich). For immunocytochemistry, cells were plated in 24-well plates on glass coverslips that had been treated with poly L-Lysine for 30 min. For the immunoblot, HEK293 cells were seeded in 6-well plates (Roth). ErCry1a (Bolte et al. 2016 (link)), erCry2a, lacking the first 39 nucleotides and erCry4a (Günther et al. 2018 (link)) were expressed as fusion proteins with GFP from the respective generated pTurbo-GFP-N vectors using 2 µl Lipofectamine 2000 reagent (Invitrogen) per µg DNA in OptiMEM medium (Life Technologies, Carlsbad, CA, USA) following the manufacturer’s protocol.

Einwich A., Seth P.K., Bartölke R., Bolte P., Feederle R., Dedek K, & Mouritsen H. (2021). Localisation of cryptochrome 2 in the avian retina. Journal of Comparative Physiology. A, Neuroethology, Sensory, Neural, and Behavioral Physiology, 208(1), 69-81.

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Culturing Primary Skin Cells

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ROC product was developed and obtained from ZOE BIO Inc. (Seoul, Korea), previously reported.
²⁵ (link) R.A. (Vitamin A acid) and AMA were purchased from Sigma (St. Louis, MO, USA). Coating Matrix (type I collagen), Fetal Bovine Serum (FBS), and Trypsin‐EDTA for primary cells were obtained from Gibco‐Life Technologies™ (Carlsbad, CA, USA). Keratinocyte Growth Medium (KGM), Hank's Balanced Salt Solution (HBSS), Antibiotics/Antimycotics, and Collagenase/Dispase were purchased from Sigma (St. Louis, MO, USA).

Trinh T.T., Choi J.H., Yang J., Kim W.H., Chien P.N., Le L.T., Ngan‐Giang N., Nga P.T., Nam S, & Heo C. (2024). Effects on keratinocytes of the traditional combination of herb extract (Royal Oji Complex) implicated the improvement of young children's skin moisture and barrier. Skin Research and Technology, 30(4), e13682.

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