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Cell culture media and supplements

Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom

Cell culture media and supplements are a range of products designed to support the growth and maintenance of cells in in vitro cell culture systems. These products provide the necessary nutrients, growth factors, and other components required for the optimal cultivation of various cell types.

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86 protocols using cell culture media and supplements

1

Chemical Sourcing for Cell Culture

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All the chemicals used in this study were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless specified otherwise. Cell culture media and supplements were obtained from Life Technologies (Grand Island, NY, USA).
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2

Culturing Human and Murine Melanoma Cell Lines

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The human melanoma A375 cell line was obtained from ATCC. Cells were grown in DMEM (Mediatech Inc.) and were supplemented with 10% fetal bovine serum (FBS), 1 µg/mL hydrocortisone, and 1% penicillin and streptomycin. The murine melanoma B78-D14 (B78) cell line is derived from B16 melanoma as previously described [13 (link)] and was obtained from Ralph Reisfeld (Scripps Research Institute) in 2002. B78 cells were grown in RPMI-1640 w/L-glutamine (Mediatech Inc.) and were supplemented with 10% FBS, 100 U/mL penicillin, and 100ug/mL streptomycin. Cell line authentication was performed per ATCC guidelines using morphology, growth curves, and Mycoplasma testing within 6 months of use. All chemicals were purchased from Sigma. Cell culture media and supplements were obtained from Life Technologies, Inc.
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3

Standardized Cell Culture Reagents

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All chemicals were purchased from Sigma Aldrich (Munich, Germany) unless stated otherwise. Cell culture media and supplements were from Life Technologies (Darmstadt, Germany) unless stated otherwise. Cell culture plastic ware was obtained from BD (Heidelberg, Germany) unless stated otherwise.
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4

Radiolabeled Thymidine Analogues for Cell Proliferation

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[methyl-3H]-3′-Deoxy-3′-fluorothymidine ([3H]-FLT; 351.5GBq/mmol), [methyl-3H]-thymidine ([3H]-TdR; 185 GBq/mmol), and [methyl-3H]- 4'-thiothymidine ([3H]-4DST; 233 GBq/mmol), were purchased from Moravek Biochemicals (Brea, CA). High-purity nitrobenzylmercaptopurine ribonucleoside (NBMPR) and other reagents were purchased from Sigma-Aldrich (St. Louis, MO). Cell culture media and supplements were purchased from Life Technologies (Carlsbad, CA).
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5

Reagents and Antibodies Used in Research

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Actinomycin D (from Streptomyces species), Rapamycin and STS were obtained from Calbiochem (Schwalbach, Germany). α-toxin from S. aureus, cycloheximide, doxorubicin and the antibody against GAPDH, β-actin and β-tubulin were purchased from Sigma-Aldrich (Deisenhofen, Germany). The antibodies against caspase-2 (ICH-1, Nedd-2), and Firefly luciferase were from Millipore (Darmstadt, Germany). Rabbit antibodies against caspase-9 and against cyclin A were from Cell Signaling (Frankfurt, Germany). Antibodies raised against HuR, anti-rabbit, anti-mouse and anti-goat horseradish peroxidase linked IgG were purchased from Santa Cruz Biotechnology (Heidelberg, Germany). Antibodies raised against phospho-S6 (Ser240/244), caspase-3 and PARP were from Cell Signaling. Protein G sepharose, the ECL system and Hyperfilm were also from GE Healthcare. Ribonucleotides and modifying enzymes were from Life Technologies (Karlsruhe, Germany). All cell culture media and supplements were purchased from Life Technologies.
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6

Estrogen Signaling Pathway Analysis

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Cell culture media and supplements were purchased from Life Technologies, Inc (Grand Island, NY). Fetal b ovine serum (FBS) was purchased from Atlanta Biologicals (Flowery Branch, GA) and dextran-coated-charcoal (DCC) stripped fetal bovine serum was purchased from Gemini Bioscience (Sacramento, CA). Estradiol (E2), dimethylsulfoxide (DMSO) and insulin were purchased from Sigma Aldrich (St. Louis, MO). Propylpyrazoletriol (PPT), diarylpropylnitrile (DPN), and G1 were purchased from Tocris Bioscience (Minneapolis, MN). Agarose was purchased from Fisher Scientific (Agawam, MA). Technovit 7100 was purchased from Heraeus Kulzer GmBH (Wehrheim, Germany).
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7

Receptor Binding Assay Protocol

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PathHunter detection reagents were from DiscoveRx (Fremont, CA). Cell culture media and supplements were from Life Technologies (Carlsbad, CA). Lance-Ultra cAMP detection reagents, Surefire ERK assay reagents, [3H]diprenorphine (DPN), and [35S]GTPγS (guanosine-5′-O-(3-thio)triphosphate) were from PerkinElmer Life Sciences (Cambridge, MA). Endomorphin I and TAN67 were obtained from Tocris. All other chemicals, unless otherwise specified, were purchased from Sigma (St. Louis, MO).
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8

Reagents and Chemicals for Cell Assays

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PathHunter detection reagents were from DiscoveRx (Fremont, CA). Cell culture media and supplements were from Life Technologies (Carlsbad, CA). All other chemicals, unless otherwise specified, were purchased from Sigma (St. Louis, MO).
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9

Transfection and Viral Infection Protocols

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Cell lines were purchased from American Type Culture Collection and cell culture media and supplements were purchased from Life Technologies, Inc., Carlsbad, CA. IMR90 primary human lung fibroblast cells were grown in Minimum Essential Media (MEM) supplemented with Earle's salt, 10% Fetal Bovine Serum (FBS) and 1-mM Sodium Pyruvate. HEK293 cells and NIH3T3 cells were each grown in Dulbecco's modified Eagle's medium supplemented with 10% FBS. Both were grown in 37°C incubator with 5% CO2.
Transfections used Lipofectamine 2000 (Life Technologies), preparing it with the DNA in Optimem (Life Technologies) per manufacturer's recommendations. The DNA–lipofectamine complexes were added to cells resuspended in growth media before plating. Transfected cells were placed back in the incubator for 3–5 h after transfection before any further treatment.
Sendai Virus (SeV) (Charles River Laboratories), was centrifuged to clarify the virus and stored at –80°C. For virus infections, we used 50 μl of virus per 1 ml of culture media. Interferon-β (Peprotech, catalog # 300–02BC) was used at 0.1 ng of IFN-β per 1 ml of culture media. These treatments were for 6 h, except for the Luciferase assays, which were for 24 h before prepping the cells for the assay.
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10

Oxidative Stress Assay Protocol

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All solvents used for chromatographic separations (ACS reagent, HPLC, and LC-MS grade) were purchased from Fisher Scientific (Fair Lawn, NJ). Bovine serum albumin (BSA), 2′,7′-dichlorodihydrofluorescin diacetate (H2DCF-DA), digitonin, DMSO, EDTA, esterase, FeSO4, flavin adenine dinucleotide phosphate (FAD), glucose-6-phosphate (G-6-P), glucose-6-phosphate dehydrogenase (G-6-P-D), H2O2, menadione, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), nicotinamide adenine dinucleotide phosphate (NADP), quercetin, L-sulforaphane, Trizma base, and Tween 20, were purchased from Sigma-Aldrich (St. Louis, MO). Cell culture media and supplements were obtained from Life Technologies, Inc. (Grand Island, NY). Deuterated NMR solvents were purchased from Cambridge Isotope Laboratories (Andover, MA).
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