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Antibiotic mixture

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The Antibiotic mixture is a laboratory product that contains a combination of antibiotics. It is used for research and development purposes in various scientific fields. The core function of this product is to provide a standardized mixture of antibiotics for experimental applications.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (13 mentions) Cell culture media, by Thermo Fisher Scientific (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) N2 supplement, by Thermo Fisher Scientific (2 mentions) B27 supplement, by Thermo Fisher Scientific (2 mentions) Accutase, by Merck Group (2 mentions) Superfrost plus slide, by Thermo Fisher Scientific (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (2 mentions) 24 well ultra low attachment plate, by Corning (2 mentions) Fetal bovine serum (fbs), by Lonza (2 mentions) Dmem medium, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Anti β actin antibody, by Merck Group (1 mentions) Mcf 7 cell line, by American Type Culture Collection (1 mentions) Cell culture reagents, by Thermo Fisher Scientific (1 mentions) Antibody against β actin, by Merck Group (1 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody, by GeneTex (1 mentions)

29 protocols using antibiotic mixture

1

Characterization of FoxQ1-overexpressing breast cancer cells

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Cell culture reagents including fetal bovine serum, cell culture media, phosphate-buffered saline (PBS), and antibiotic mixture were purchased from Life Technologies-Thermo Fisher Scientific (Waltham, MA). An antibody against FoxQ1 was from Santa Cruz Biotechnology (Dallas, TX). An antibody specific for detection of phospho-(Ser10) histone H3 was from Cell Signaling Technology (Danvers, MA). Anti-β-Actin antibody was from Sigma-Aldrich (St. Louis, MO). The MCF-7 cell line was purchased from the American Type Culture Collection and authenticated by us in 2015 and 2017. The SUM159 cell line was purchased from Asterand (Detroit, MI) and authenticated by us in 2015 and 2017. Details of stable transfection of MCF-7 and SUM159 cells with pCMV6 empty vector and the same vector encoding FoxQ1 and their culture conditions have been described by us previously.10 (link) The HMLE cells stably transfected with FoxQ1 and EV cells were generously provided by Dr. Guojun Wu (Karmanos Cancer Institute, Departments of Oncology and Pathology, Wayne State University, Detroit, MI) and maintained as recommended by the provider.7 (link)
Kim S.H., Hahm E.R., Singh K.B, & Singh S.V. (2020). Novel mechanistic targets of forkhead box Q1 transcription factor in human breast cancer cells. Molecular carcinogenesis, 59(10), 1116-1128.
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2

Lipid Metabolism Pathway Assays

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LLM (purity ≥ 98%) was purchased from the Cayman Chemical Company. Fetal bovine serum, antibiotic mixture, and other cell culture reagents were purchased from Life Technologies-ThermoFisher Scientific. The RPMI 1640 media was purchased from Mediatech. Anti-ACC1 (cat. #4190) and anti-FASN (cat. #3180) antibodies were purchased from Cell Signaling Technology. The anti-ACLY (cat. #ab40793) and anti-SREBP1 (cat. #ab28481) antibodies were purchased from Abcam. Kits for determination of total free fatty acids (cat. #MAK044) and anti-ACC1 antibody for immunohistochemistry (cat. #SAB4501396) were purchased from Sigma-Aldrich.
Singh K.B., Hahm E.R., Pore S.K, & Singh S.V. (2019). Leelamine is a Novel Lipogenesis Inhibitor in Prostate Cancer Cells In Vitro and In Vivo. Molecular cancer therapeutics, 18(10), 1800-1810.
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3

FoxQ1 Expression in Breast Cancer Cells

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The WA (purity > 95%) was purchased from ChromaDex (Irvine, CA) and dissolved in dimethyl sulfoxide (DMSO). Reagents for cell culture, including fetal bovine serum, cell culture media, and antibiotic mixture were purchased from Life Technologies-Thermo Fisher Scientific (Waltham, MA). An antibody against FoxQ1 was purchased from Proteintech (Rosemont, IL) and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody was from Genetex (Irvine, CA). An antibody against β-Actin was from Sigma-Aldrich (St. Louis, MO). The MDA-MB-231 and MCF-7 cell lines were purchased from the American Type Culture Collection (Manassas, VA), whereas SUM159 cell line was from Asterand Bioscience (Detroit, MI). Each cell line was last authenticated by us in 2017 by short tandem repeat profiling. Each cell line was cultured according to the supplier’s recommendations. Details of stable transfection of MCF-7 and SUM159 cells with pCMV6 empty vector and the same vector encoding FoxQ1 have been described by us previously (24 (link)).
Kim S.H., Singh K.B., Hahm E.R, & Singh S.V. (2021). The Role of Forkhead Box Q1 Transcription Factor in Anticancer Effects of Withaferin A in Breast Cancer. Cancer prevention research (Philadelphia, Pa.), 14(4), 421-432.
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4

Establishment of Gastric Cancer Stemness and Resistance

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Gastric epithelial cells (GES-1) and GC cell lines (AGS, SCG-7901, MGC-803, SNU-16, and MKN1) were purchased from ATCC and stored in RPMI 1640 medium containing 10% FBS and antibiotics. Cell microsphere culture to obtain stemness was conducted with reference to Liu et al. (25 (link)). In brief, AGS cells (100 cells/well) were placed in serum-free RPMI-1640 medium supplemented with 1% N-2 supplement, 2% B-27 supplement (Invitrogen, Carlsbad, CA, USA) (Invitrogen), 1% antibiotic mixture (Invitrogen, Carlsbad, CA, USA), 20 ng/mL human FGF-2, and 100 ng/mL EGF (Chemicon). After 2 weeks, spheroid formation was analyzed and observed with an inverted microscope (Olym-pus, Tokyo, Japan) (Olympus) 40× magnification. Based on the method of Jiang et al. (26 (link)), the AGS cells were continuously exposed to cisplatin (from 0.05 to 1 mg/mL) to establish a cisplatin-resistant AGS cell (AGS/DDP) line.
Ma Y., Xue H., Wang W., Yuan Y, & Liang F. (2020). The miR-567/RPL15/TGF-β/Smad axis inhibits the stem-like properties and chemo-resistance of gastric cancer cells. Translational Cancer Research, 9(5), 3539-3549.
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5

Isolation and Culture of Rat Neurons

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Adult male spontaneously hypertensive rats (250-350g) were purchased from Charles River Laboratories. Neurobasal medium, B27 (without retinoic acid), recombinant human FGF-2 and antibiotic mixture were purchased from Invitrogen (Carlsbad, CA, USA). Other reagents were purchased from the following suppliers: CellTiter-Glow reagent (Promega Inc. Madison, WI, USA); Cytochrome C Oxidase kit, Accutase, Heparin and Glutamine were from Sigma chemical company (St. Louis MO, USA).
Kalluri H.S, & Dempsey R.J. (2014). D609 mediated inhibition of ATP synthesis in neural progenitor cells. Neuroreport, 25(10), 777-781.
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6

Immunohistochemistry of Lymphatic and Vascular Markers

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Dulbecco’s phosphate-buffered saline (DPBS) (Catalog no. 14040-133, Gibco by Life Technologies Corporation, Grand Island, NY 14072, USA)
D-MEM/F12 (Catalogue number 11039-021, Gibco by Life Technologies Corporation, Grand Island, NY 14072, USA) supplemented with antibiotic mixture (Invitrogen) to achieve a final concentration of 100 units of penicillin/100 μg of streptomycin/mL
Ketamine/xylazine (Catalogue number: K113–10ML, Sigma-Aldrich, St. Louis, MO 63178, USA)
1XProtease inhibitor cocktail (Catalog Number P8340, Sigma-Aldrich)
Fentanyl (Catalogue number F3886, Sigma-Aldrich) / Droperidol (Catalogue Number D1414, Sigma-Aldrich)
Diazepam (Catalogue number NDC0409-3213-12, Hospira Inc., Lake Forest, IL 60045, USA)
ovalbumin-conjugated Alexa Fluor 647 (Thermo Fisher Scientific O34784)
aCSF (Harvard Apparatus 597316)
Superfrost Plus Slides (Thermo Fisher Scientific)
Fisherbrand Cover Glasses 50X22 No 1 (Fisher 12-545E)
Alexa Flour 488-conjugated rat anti-mouse Lyve1 (Invitrogen 14-0443-82, clone ALY7)
Armenian hamster anti-mouse CD31 (Millipore Sigma MAB1398Z, clone 2H8)
Alexa Fluor 594-conjugated goat anti-Armenian hamster (Jackson ImmunoResearch, 127-585-160)
DAPI (Sigma-Aldrich D9542)
ProLong Gold Antifade Mountant (Invitrogen)
Zawieja D.C., Thangaswamy S., Wang W., Furtado R., Clement C.C., Papadopoulos Z., Vigano M., Bridenbaugh E.A., Zolla L., Gashev A.A., Kipnis J., Lauvau G, & Santambrogio L. (2019). Lymphatic Cannulation for Lymph Sampling and Molecular Delivery. Journal of immunology (Baltimore, Md. : 1950), 203(8), 2339-2350.
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7

ESCC Cell Line Cultivation Protocol

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Three human ESCC cell lines, KYSE510 and KYSE150, were used in this study. They were maintained in RPMI 1640 supplemented with 10% fetal bovine serum and 1 × antibiotic mixture (Invitrogen, Carlsbad, CA, USA). All cells were cultured at 37°C in a humidified incubator containing 5% CO2.
Wang H., Zhang Y., Yun H., Chen S., Chen Y, & Liu Z. (2017). ERK expression and its correlation with STAT1 in esophageal squamous cell carcinoma. Oncotarget, 8(28), 45249-45258.
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8

Generating Tumorspheres from HCT116 Cells

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Tumorspheres were generated by seeding the HCT116 adherent cells into serum-free DMEM culture medium containing, B27 (1x, Gibco), EGF (20 ng/ml, Peprotech), bFGF (10 ng/ml, Peprotech), 1% antibiotic mixture (invitrogen), and the healthy adherent cells were plated in 100-mm ultra-low attachment plate (Corning) at 1 × 106 cells per plate in humidified incubator at 37°C in 5% CO2. After 7 days, the primary tumorspheres were counted and dissociated at the density of 1,000 cells per milliliter and 500 mL single cell suspension was seeded in each well of 24-well ultra-low attachment plate (Corning) in serum-free medium described above. After 7 days later, secondary tumorsphere were counted and took picture under inverted microscopy.
Kangwan N., Kim Y.J., Han Y.M., Jeong M., Park J.M., Go E.J, & Hahm K.B. (2015). Sonic hedgehog inhibitors prevent colitis-associated cancer via orchestrated mechanisms of IL-6/gp130 inhibition, 15-PGDH induction, Bcl-2 abrogation, and tumorsphere inhibition. Oncotarget, 7(7), 7667-7682.
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9

Transfection of HeLa cells for microscopy

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HeLa cells were grown at 37 °C/5% CO2 in Dublecco’s modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (Gibco) and 1% antibiotic mixture (Invitrogen Corporation Pontoise). HeLa cells were transfected with a mixture of pNL4.3-MS2-Δenv plasmid or derivatives with MCP-eGFP-NLS plasmid (ratio 0.6/0.4) by incubating cells with 2 μg of DNA plasmids for confocal microscopy or 10 μg of plasmid for TEM with jetPEI (Life Technologies).
Durand S., Seigneuret F., Burlaud-Gaillard J., Lemoine R., Tassi M.F., Moreau A., Mougel M., Roingeard P., Tauber C, & de Rocquigny H. (2021). Quantitative analysis of the formation of nucleoprotein complexes between HIV-1 Gag protein and genomic RNA using transmission electron microscopy. The Journal of Biological Chemistry, 298(1), 101500.
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10

ESCC Cell Line Maintenance and Clinical Sample Collection

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Human ESCC cell lines, EC1 and KYSE 150, were used in this study. Cells were maintained in RPMI 1640 (KYSE150), or Dulbecco’s modified Eagle’s medium supplemented (EC1) with 10% fetal bovine serum, and 1 × antibiotic mixture (Invitrogen, Carlsbad, CA, USA). All cells were cultured at 37 °C in a humidified incubator containing 5% CO2.
We randomly collected 201 consecutive ESCC samples at the Shantou Tumor Hospital between 2005 and 2012. All patients underwent potentially curative surgery without preoperative chemotherapy or radiotherapy. In this cohort, 150 were men and 51 were women; the range of ages was 36–81 years, with a median of 57 years. Follow-up data were available for 130 patients; most (113, 86.9%) died during the follow-up period (median survival, 21.5 months). Of the 201 ESCC tumors, 33 case-matched normal esophageal tissues adjacent to the tumors were included in the study. Written informed consents were obtained from the patients, and the study was reviewed and approved by the institutional ethics committee of Shantou University Medical College.
Zhang Y., Chen Y., Yun H., Liu Z., Su M, & Lai R. (2017). STAT1β enhances STAT1 function by protecting STAT1α from degradation in esophageal squamous cell carcinoma. Cell Death & Disease, 8(10), e3077-.
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