Human lung cancer cells (A549, H1299, H460, and H1650) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (Welgene, Gyeongsangbuk-do, Korea) supplemented with 10% fetal bovine serum (Welgene) and 100 units/mL penicillin–streptomycin solution (Gibco, Grand Island, NY, USA) at 37 °C in a humidified 5% CO2 atmosphere.
1640 medium
Manufactured by Welgene
Sourced in United States
1640 medium is a cell culture medium formulated to support the growth and maintenance of a variety of cell lines. It provides the necessary nutrients and supplements for cell proliferation and viability. The composition of 1640 medium is designed to create an optimal environment for cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Welgene (5 mentions)
Fetal bovine serum (fbs), by Welgene (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dulbecco s modified eagle s medium, by Welgene (4 mentions)
Streptomycin, by Welgene (4 mentions)
Penicillin streptomycin, by Welgene (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Antibiotic solution, by Welgene (2 mentions)
H1299, by American Type Culture Collection (1 mentions)
H1650, by American Type Culture Collection (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Mhy1485, by Merck Group (1 mentions)
Sodium pyruvate, by Welgene (1 mentions)
Bradford reagent, by Bio-Rad (1 mentions)
Pcr primer, by Bioneer (1 mentions)
Enzyme linked chemiluminescence ecl western detection reagents, by Thermo Fisher Scientific (1 mentions)
Cell culture plastic wares, by SPL Life Sciences (1 mentions)
Control sirna sc 37007, by Santa Cruz Biotechnology (1 mentions)
20 protocols using 1640 medium
1
Culturing Human Lung Cancer Cells
2
Culturing HepG2 Cells for Toxicological Studies
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The human hepatic carcinoma cell line HepG2 was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). Cells with a passage number between 11 and 15 were used and cultured in a 150 mm2 cell culture dish. Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Welgene, Gyeongsan, Republic of Korea) supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA), 100 μg/mL penicillin/streptomycin (Welgene, Gyeongsan, Republic of Korea), and 1 mM sodium pyruvate (Welgene, Gyeongsan, Republic of Korea) in a humidified incubator at 37 °C with 5% CO2. The culture medium was changed every 3 days. For the assays, cells were treated with or without B[a]P and transfected with CYP1A1, CYP1B1, or control siRNA for 24 h or 48 h. A 10 mM stock solution of B[a]P in DMSO was diluted to a final concentration of 10 μM in cell culture media. Moreover, in the case of MHY 1485 (Sigma-Aldrich Chemical, St. Louis, MO, USA), an mTOR activator, it was additionally treated at a concentration of 2 μM during the transfection process and cultured for 24 h or 48 h.
3
Dasatinib's Anticancer Effects Elucidated
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Dasatinib was purchased from LC Laboratories. Roswell Park Memorial Institute (RPMI)‐1640 medium, Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS), penicillin‐streptomycin and phosphate‐buffered saline (PBS) were bought from WelGENE. 3‐(4,5‐dimethylthiazol‐2‐y)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS) reagent was from Promega. Bradford reagent was from Bio‐Rad. Protease inhibitor cocktail (PIC, 100×) and z‐VAD‐fmk (627610) were purchased from Calbiochem. Control siRNA (sc‐37007) and Src (sc‐29228) were obtained from Santa Cruz Biotechnology. Enzyme‐linked chemiluminescence (ECL) Western detection reagents were bought from Thermo Scientific. PCR primers were purchased from Bioneer. LY294002 and PD98059 were from Biomol Research Lab. Cell culture plastic wares were purchased from SPL Life Sciences. A detailed list of antibodies used in this study is included in Supplementary Table S1 .
4
Cytotoxicity Assay and Cell Signaling Analysis
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Roswell Park Memorial Institute (RPMI) 1640 Medium, Dulbecco's Modified Eagle's Medium (DMEM), penicillin/streptomycin, amphotericin B, and Trypsin-EDTA were purchased from Welgene (Seoul, Republic of Korea). Fetal bovine serum (FBS) was purchased from J R Scientific (Woodland, CA, USA). CuD was purchased from Extrasynthese (Genay, France). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), 7-aminoactinomycin D (7-AAD), 2′,7′-dichlorofluorescein diacetate (DCF-DA), and N-acetyl-L-cysteine (NAC), SP600125 and SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA). Annexin V was purchased from BD Biosciences (Franklin Lakes, NJ, USA). Primary antibodies against phospho-cdc2, phospho-25c, p21, cleaved caspase-7 and -8, cleaved PARP, JNK, c-jun, phospho-c-jun, p38, phospho-p38, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA). Primary antibodies against cyclin B1, cdc2, cdc25c, and phospho-JNK were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The secondary antibody, horse antimouse immunoglobulin G (IgG)-horseradish peroxidase (HRP) and goat antirabbit IgG-HRP were purchased from Cell Signaling Technology.
5
Xenograft model of renal cancer
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The human renal cancer cell lines A498 and Caki-1 were obtained from the Korean Cell Line Bank. A498 and Caki-1 cells were cultured in Roswell Park Memorial Institute-1640 medium (WELGENE, Daegu, Republic of Korea) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Carlsbad, CA, USA) and 1% penicillin–streptomycin (Gibco). The cells were maintained in an incubator conditioned with 5% CO2 at 37 °C. Four-week-old female BALB/c nu/nu mice were purchased from ORIENT BIO. Animals were maintained at 23 ± 1 °C and 50 ± 10% humidity under specific pathogen-free conditions. The light–dark period was cycled every 12 h, and food and water were provided ad libitum. Animal experimental procedures were reviewed and approved by the CHA University Animal Care and Use Committee (IACUC210152).
6
Established Cell Lines and Antibodies
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The human TNBC cell line (MDA-MB-231) and murine macrophage (Raw264.7) were obtained from the Korean Cell Line Bank (Seoul, Korea). All cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (WelGENE, Daegu, Korea) containing 10% fetal bovine serum (FBS) and supplemented with a 1% antibiotic solution containing penicillin and streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). Cells were cultured in a 5% CO2 incubator at 37°C. The primary antibodies used in this study were anti-cytokeratin 8/18/19 antibody, anti-GFP antibody, anti-RFP antibody, anti-CD63 antibody, anti-ALIX antibody, and anti-calnexin antibody, purchased from Abcam (Cambridge, MA, USA). Anti-NOS2 antibody, anti-CD206 antibody, anti-arginase-1 antibody, and anti-GAPDH antibody were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Anti-β-actin antibody was purchased from Sigma (St. Louis, MO, USA).
7
Culturing NSCLC and NK Cell Lines
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NCI-H23 (Korean Cell Line Bank, Seoul, Korea), a human NSCLC cell line, was maintained in Roswell-Park-Memorial-Institute (RPMI) 1640 medium (WELGENE, Gyeongsan, Korea) with fetal bovine serum (10%; WELGENE) and penicillin (1%; WELGENE). The NK92 cell line (American Type Culture Collection, Rockville, MD), a human NK cell line, was maintained in the α-minimum essential medium with fetal bovine serum (12.5%), horse serum (12.5%), recombinant human interleukin-2 (200 U/mL), 2-mercaptoethanol (0.1 mM), and l-glutamine (2 mM). The cells were incubated at 37°C humidified air containing 5% CO2.
8
Anti-inflammatory Mechanisms in Immune Cells
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Fetal Bovine Serum (FBS), Dulbecco's Modified Eagle Medium (DMEM), Roswell Park Memorial Institute (RPMI) 1640 Medium, streptomycin, and penicillin were from Welgene (Daegu, Republic of Korea). The oligonucleotide primers used, shown in Table 1 , were purchased from Bioneer (Daejeon, Republic of Korea). The TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA). 2′, 7′–dichlorofluorescin diacetate (DCFDA), lipopolysaccharide (LPS), Bay-11, N-acetyl-cysteine (NAC), phorbol 12-myristate 13-acetate (PMA), and DAPI stain were obtained from Sigma–Aldrich (St. Louis, MO, USA). Antibodies for western blot analysis, inducible NO synthase (iNOS; #2982), cyclooxygenase-2 (COX-2; #4842), p-TAK1 (phospho-transforming growth factor beta-activated kinase 1; #4531), p-IκBα (#2859), p-NFκB p65 (#3033), T-NFκB p65 (#8242), β-Actin (#4967), total-extracellular-signal-regulated kinase (T-ERK; #9102), p-ERK (#9101), total-c-Jun N-terminal kinase (T-JNK; #9252), p-JNK (#9251), T-p38 (#9212), p-p38 (#9211), poly (ADP ribose) polymerase (PARP; #9542), HRP-linked antibody (#7074), and ALexa Fluor 555® conjugated secondary antibody (#4413) were from Cell Signaling Technology (Danvers, MA, USA).
9
Epithelial Cell Adherence Assay
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The human ileocecal epithelial cell line HCT-8 and the human bladder epithelial cell line T-24 were provided by the Korean Cell Line Bank (Seoul, Korea). Cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (Welgene, Gyeongsan, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 mg/mL). For epithelial cell adherence assays, HCT-8 and T-24 cells were seeded at ~104 cells/well in a 96-well plate (polystyrene; clear flat bottoms; Corning, NY, USA) and grown in 5% CO2 for 24 h at 37 °C.
10
Generation of Bone Marrow-Derived Immune Cells
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Bone marrow-derived dendritic cells (BMDCs) and bone marrow-derived macrophages (BMDMs) were generated by flushing bone marrow cells from femurs and tibias. BMDCs were cultured for 6 days in Roswell Park Memorial Institute (RPMI) 1640 medium (Welgene Co., Daegu, Korea) supplemented with 10% fetal bovine serum (FBS) (Welgene), 100 unit/mL of penicillin/100 μg/mL of streptomycin (Welgene), 0.1 mM nonessential amino acids (Lonza, Basel, Switzerland), 50 μM β-mercaptoethanol (Lonza), 1 mM sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA), 20 ng/mL GM-CSF (CreaGene, Gyeonggi, Korea), and 10 ng/mL IL-4 (CreaGene). BMDMs were cultured for 6 days in Dulbecco’s modified eagle’s medium (Welgene) containing 10% FBS, 50 ng/mL mouse macrophage colony stimulating factor (M-CSF) (R&D System, Minneapolis, MN, USA), and 100 unit/mL of penicillin/100 μg/mL of streptomycin. Cultured cells were incubated at 37 °C in a 5% CO2 atmosphere.
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