The murine melanoma cell line SM1 (a BRAFV600E-driven melanoma) was obtained from the laboratory of Dr. A. Ribas at the University of California Los Angeles [36 (link)]. The murine melanoma cell line B16F10 was purchased from ATCC (Manassas, VA, USA). Cell culture media Dulbecco’s modified Eagle Medium (DMEM) was purchased from Thermo Fisher Scientific and Hyclone RPMI 1640 media was purchased from GE Healthcare Life Sciences (Pittsburgh, PA, USA). When needed, cell culture media was supplemented with 1% penicillin/streptomycin (PenStrep) purchased from Corning (Corning, NY, USA). Fetal bovine serum (FBS) was obtained from Thermo Fisher Scientific.
Hyclone rpmi 1640 media
Manufactured by GE Healthcare
Sourced in United States
Hyclone RPMI 1640 media is a powdered cell culture medium formulation developed for the in vitro cultivation of a variety of mammalian cells. It provides a balanced salt solution and a source of energy, vitamins, and amino acids to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Sodium pyruvate, by GE Healthcare (2 mentions)
Non essential amino acid (neaa), by Lonza (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin pen strep, by Corning (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Endothelial cell growth medium, by PELOBiotech (1 mentions)
Penicillin, by PAN Biotech (1 mentions)
Amphotericin b, by PAN Biotech (1 mentions)
Streptomycin, by PAN Biotech (1 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Docetaxel, by Yuanye Bio-Technology (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by GE Healthcare (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
L glutamine, by Lonza (1 mentions)
7 protocols using hyclone rpmi 1640 media
1
Murine Melanoma Cell Line Protocols
2
Cultivation of THP-1 and HUVEC Cells
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THP-1 cells (ACC-16) were obtained from the Leibniz Institute DMSZ—German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany) and were cultured in HyClone™ RPMI 1640 media (GE Healthcare, Chicago, IL, USA) containing 10% fetal bovine serum (FBS, Biochrom, Berlin, Germany), 100 U/mL penicillin (PAA, Pasching, Austria) and 100 µg/mL streptomycin (PAA) at 37 °C in an atmosphere with 5% CO2. Primary human umbilical vein endothelial cells (HUVECs) were obtained from Provitro (Berlin, Germany) and cultured in Endothelial Cell Growth Medium (PELOBiotech, Planegg, Germany) containing 10% FBS, 100 U/mL penicillin, 100 µg/mL streptomycin and 2.5 µg/mL amphotericin B (PAN-Biotech, Aidenbach, Germany). HUVECs were cultivated until passage 2 and used for experiments at passage 3. Both cell types were cultured at 37 °C in an atmosphere with 5% CO2.
3
Evaluating Docetaxel, Pemetrexed, and Cisplatin in Lung Cancer
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The materials used in the present study were as follows: Docetaxel (Shanghai Yuanye Bio-Technology Co., Ltd., Shanghai, China); pemetrexed (National Institutes for Food and Drug Control, Beijing, China); cisplatin (National Institutes for Food and Drug Control, Beijing, China); the A549 cell line (JRDUN Biotechnology, Shanghai, China); Hyclone RPMI 1640 media (GE Healthcare; Logan, UT, USA); fetal bovine serum (FBS; MRC, Jintan, Jiangsu, China); 0.25% EDTA-trypsin (Beijing Leagene Biotechnology Co., Ltd., Beijing, China); CellTiter 96 Aqueous One Solution Cell Proliferation assay (Promega Corporation, Madison, WI, USA); TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA); RevertAid First Strand cDNA Synthesis kit (Thermo Fisher Scientific, Inc.); Real Time PCR Easy-SYBR Green I (Foregene Co., Ltd., Chengdu, Sichuan, China); primers (Shanghai Generay Biotech Co., Ltd., Shanghai, China); polyvinylidene difluoride (PVDF) membrane (EMD Millipore, Billerica, MA, USA); BCIP/NBT alkaline phosphatase color development kit (Beijing Huamaike Biotechnology, Beijing, China); and anti-ERCC1 (human) antibody, anti-β-actin (human) antibody and alkaline phosphatase-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) antibody (Proteintech Group Inc., Rosemont, IL, USA).
4
In Vitro Transfection of L929 Cells
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For in vitro transfection of L929 cells, mRNAs (MmCyp7b1-HA, HsCYP7B1-HA) were complexed with Lipofectamine2000 (Life Technologies) at 1.5 μL per μg of mRNA and transfected into cells according to manufacturer’s instructions. For western blot analysis, 3 × 105 L929 cells were seeded 1 day before transfection in 6-well plates and transfected with 2 μg of complexed mRNA for each construct. For immunocytochemical staining, 5 × 104 L929 cells were seeded on 24-well plates on coverslips (Roth) 1 day before transfection with 1 μg mRNA as described above. Approximately 2 h after transfection, the transfection mix was replaced with HyClone RPMI 1640 media (GE Healthcare) supplemented with 2 mM L-glutamine (Lonza), 100 U/mL penicillin (Lonza), 100 U/mL streptomycin (Lonza), and 10% fetal calf serum (GE Healthcare).
5
Cigarette Smoke Extract Generation
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The water-soluble components of tobacco smoke were generated in the form of cigarette smoke extract (CSE) as described in detail previously.19 (link),20 (link) Briefly, CSE was generated by leading mainstream smoke from ten medium nicotine and tar commercial cigarettes (Marlboro™, Philip Morris Int, Richmond, VA, USA) through 15 mL of cell culture medium (HyClone RPMI1640 Media, GE Healthcare Bio-Sciences AB) by vacuum suction at room temperature. The burning time for each cigarette was 3 minutes. The prepared solution with CSE was thereafter passed through a sterile filter and divided into aliquots and stored frozen (−80°C). The chosen concentration of CSE (1:25) resulted in IL-16 response with well-preserved cell viability (see “Results” section).
6
Culturing Human Lymphoblastoid TK6 Cells
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Human lymphoblastoid TK6 cells were purchased from MilliporeSigma (cat. 95111735) and grown in HyClone RPMI-1640 media (SH30027.01; GE Healthcare Life Sciences, Utah, USA) supplemented with 10% fetal bovine serum (SH30071.03GE; Healthcare Life Sciences), 1% non-essential amino acid (13-114E; Lonza, NJ, USA), 1% sodium pyruvate (SH30239.01; GE Healthcare Life Sciences) and 1% penicillin-streptomycin (15070063; Gibco, Thermo Fisher Scientific, MA, USA). Cells were grown at 37 °C in a humidified atmosphere of 5% CO2 in air and were routinely passaged to ensure they remained in the exponential growth phase. Cells never exceeded passage 30 and their average doubling time was 14–15 h.
7
Growth and Maintenance of TK6 Human Lymphoblastoid Cells
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Human lymphoblastoid TK6 cells were purchased from MilliporeSigma (MilliporeSigma, Billerica, MA, USA; cat. 95111735) and were grown in HyClone RPMI‐1640 media (GE Healthcare Life Sciences, Logan, Utah, USA; cat. SH30027.01) supplemented with 10% fetal bovine serum (GE Healthcare Life Sciences, Logan, UT; cat. SH30071.03), 1% nonessential amino acid (Lonza, Allendale, NJ, USA; cat. 13–114E), 1% sodium pyruvate (GE Healthcare Life Sciences, Logan, UT; cat. SH30239.01), and 1% penicillin–streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA; cat. 15070063). The TK6 cells were routinely passaged to ensure that they were kept in the exponential growth phase (1 × 104–1.5 × 106 cells/ml) for all experiments and their doubling time was approximately 15 h.
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