MDA-MB-453, UACC-893, HCC-2218, HCC-1419 (ATCC, Manassas, VA, USA), and ZR-75-1 cells (Institute of Development, Aging and Cancer (IDAC), Miyagi, Japan) were cultured in Roswell Park Memorial Institute medium (RPMI-1640, FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS, NICHIREI BIOSCIENCES INC., Tokyo, Japan), 100 U/mL penicillin (Meiji-Seika Pharma Co., Ltd., Tokyo, Japan), and 100 µg/mL streptomycin (Meiji-Seika Pharma) at 37 °C with 5% CO2. MDA-MB-361 and HCC-202 cells (ATCC) were cultured in RPMI-1640 (FUJIFILM Wako Pure Chemical Corporation) supplemented with 15% heat-inactivated FBS, 100 U/mL penicillin (Meiji Seika Pharma), and 100 µg/mL streptomycin (Meiji-Seika Pharma) at 37°C with 5% CO2. BT-474, UACC-812, and 293T cells (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% heat-inactivated FBS, 100 U/mL penicillin (Meiji Seika Pharma), and 100 µg/mL streptomycin (Meiji-Seika Pharma) at 37 °C with 5% CO2. Luc2-expressing breast cancer cell lines were established by infection with lentivirus (pLenti-PEF1-luc2-IRES-BlaR) and selection by blasticidin (Kaken Pharmaceuticals Co. Ltd., Tokyo, Japan).
Penicillin
Manufactured by Meiji Seika Pharma
Sourced in Japan, United States
Penicillin is a laboratory equipment used in the production and purification of the antibiotic drug penicillin. It is a critical component in the pharmaceutical manufacturing process.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Meiji Seika Pharma (44 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Nichirei Biosciences (4 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (4 mentions)
Rpmi 1640, by Fujifilm (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Sodium pyruvate, by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (3 mentions)
Rpmi 1640, by Nissui Pharmaceutical (3 mentions)
Penicillin, by Fujifilm (3 mentions)
Streptomycin, by Fujifilm (3 mentions)
Insulin, by Merck Group (3 mentions)
Fetal calf serum (fcs), by Merck Group (2 mentions)
Penicillin, by Nissui Pharmaceutical (2 mentions)
Streptomycin, by Nissui Pharmaceutical (2 mentions)
Superfect transfection reagent, by Qiagen (2 mentions)
Pegfp f, by Takara Bio (2 mentions)
Fetal bovine serum (fbs), by Biowest (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
52 protocols using penicillin
1
Culturing Breast Cancer Cell Lines
2
Cell Culture Conditions for HeLa, HEK293, and U87-MG
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HeLa and HEK293 cells (ATCC) were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% fetal calf serum (Merck KGaA), 2 mM L-glutamine (Thermo Fisher Scientific), 100 U/ml penicillin (Meiji Seika Pharma Co, Ltd), and 100 μg/ml streptomycin (Meiji Seika Pharma Co, Ltd) at 37 °C in 5% CO2. U87-MG cells (provided by Dr Tsuneo Imanaka (University of Toyama)) were cultured in Eagle’s Minimum Essential medium (Nissui) supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in 5% CO2. Before drug treatment, the culture medium of HeLa cells was changed to DMEM with 0.5% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin, and then cells were incubated at 37 °C in 5% CO2 for 16 to 24 h.
3
Culturing and Transfecting Human Cell Lines
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Human embryonic kidney cell lines (HEK293, HEK293T) and HepG2 were cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma) containing 10% fetal bovine serum (FBS), 30 U/ml penicillin, and 30 μg/ml streptomycin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan). Human lung fibroblast (WI-38) cells were cultured in modified Eagle's medium (MEM) containing 10% FBS, 30 U/ml penicillin, and 30 μg/ml streptomycin. All cells were grown at 37°C in a humidified atmosphere of 95% air, 5% CO2. HepG2 (RCB1886) and WI-38 (RCB0702) cells were purchased from RIKEN BRC (Tsukuba, Japan).
HEK293 cells were co-transfected with the recombinant plasmids and pEGFP-F (Clontech Laboratories, Palo Alto, CA, USA) as a transfection marker using SuperFect Transfection Reagent (QIAGEN, Valencia, CA, USA) according to the manufacturer's instructions. Transfected cells were grown for 36–40 h prior to performing [Ca2+]i measurements. HEK293T cells were transfected with the recombinant plasmids using Lipofectamine 2000 transfection reagent (Invitrogen, Life Technologies Corporation, Grand Island, NY, USA) according to the manufacturer's instructions and the transfected HEK293T cells were grown for 36 h prior to performing in situ hybridization.
HEK293 cells were co-transfected with the recombinant plasmids and pEGFP-F (Clontech Laboratories, Palo Alto, CA, USA) as a transfection marker using SuperFect Transfection Reagent (QIAGEN, Valencia, CA, USA) according to the manufacturer's instructions. Transfected cells were grown for 36–40 h prior to performing [Ca2+]i measurements. HEK293T cells were transfected with the recombinant plasmids using Lipofectamine 2000 transfection reagent (Invitrogen, Life Technologies Corporation, Grand Island, NY, USA) according to the manufacturer's instructions and the transfected HEK293T cells were grown for 36 h prior to performing in situ hybridization.
4
Establishing Mouse Tumor Cell Lines
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The E0771 mouse breast cancer and the mouse 3LL cell lines were purchased from CH3 BioSystems and JCRB Cell Bank, respectively. The MC-38 mouse colon adenocarcinoma cell line was purchased from Kerafast. The cells were cultured in RPMI 1640 medium (Nacalai Tesque) supplemented with 10% fetal bovine serum (Biowest), penicillin (100 U/ml; Meiji Seika Pharma), and streptomycin (100 µg/ml; Meiji Seika Pharma). To establish GFP- and OVA-expressing E0771 cell lines (E0771-GFP and E0771-OVA, respectively), E0771 cells were transfected with pcDNA3-EGFP or pcDNA3-OVA plasmids (Addgene) and Lipofectamine LTX Reagent with Plus Reagent (Thermo Fisher Scientific) and selected with G418 (Wako) for the expression of each transgene. C57BL/6J WT, B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J (C57BL/6J-tdTomato), and C57BL/6J-Tmem173gt/J (STINGΔ) mice were obtained from Charles River Japan or The Jackson Laboratory. C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I) mice were obtained from The Jackson Laboratory. C57BL/6-Ly5.1 mice (RBCC00144) were provided by the RIKEN BRC through the National BioResource Project of the MEXT/AMED, Japan. Mice were maintained and handled in accordance with the animal facility at Asahikawa Medical University or Hokkaido University.
5
Murine Dendritic Cell IL-12 Production
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Female C3H/HeN mice were sacrificed by cervical dislocation and bone marrow cells were collected. The bone marrow cells were cultured in RPMI1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (Hyclone Laboratories), 200 U/mL penicillin (Meiji Seika Pharma, Tokyo, Japan), 200 µg/mL streptomycin (Meiji Seika Pharma) and 20 ng/mL murine GM-CSF (PeproTech, NJ, U.S.A.) at 37 °C in 5% CO2/95% air. On the second day of culture, 75% of the culture medium was replaced, and on the fourth day, all of the medium was replaced. On the seventh day, cells adhering to the dish were collected with a scraper. The cell suspension (2 × 105/100 µL/well) was added to 96-well plates and incubated with 20 ng/mL murine GM-CSF for 30 min. Thereafter, the culture supernatant was removed and each stimulant was added to the above medium containing 0.3 ng/mL LPS without GM-CSF. After 24 h, the culture supernatant was collected and IL-12p40 production was measured using an ELISA kit (R&D Systems, MN, U.S.A.) according to the manufacturer’s instructions.
6
Calcium-Induced Keratinocyte Differentiation
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Normal human epidermal keratinocytes (NHEKs) (FC-0025, Lifeline Cell Technology, Frederick, USA; purchased from Kurabo, Osaka, Japan) were cultured in Humedia-KG2 medium supplemented with growth factors (Kurabo). The cells were exposed to 2 mM CaCl2 for 2 days to induce differentiation. The human keratinocyte cell line HaCaT (gift from Dr. Manabe) has been maintained for more than ten years in DMEM/Ham’s F12 (DH) medium (Fujifilm-Wako, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (CELLect, MP Biomedicals, Illkirch, France), 50 U/mL penicillin, and 50 μg/mL streptomycin (Meiji Seika Pharma, Osaka, Japan) (DH10). These cells, their transgenic derivatives, and Cos7 cells were cultured in DH10 medium. For certain cultures, the cells were treated with 5 μM MG132 (Abcam, Cambridge, UK) for 4 or 7 h. For induction of tight junction (TJ) formation, cells were treated with 10 mM CaCl2 and 40 μM the c-Jun N-terminal kinase (JNK) inhibitor SP600125 (Sigma‒Aldrich, St. Louis, USA) for 24 h.
7
Culturing Primary Human Retinal Endothelial Cells
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Primary human retinal microvascular endothelial cells (HRMECs) were obtained from DS Pharma Biomedical (Osaka, Japan). HRMECs were maintained in a humidified atmosphere with 5.0% CO 2 at 378C, with Cell Systems Corporation (CSC) complete medium (DS Pharma Biomedical) including 100 U/mL penicillin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan) and 100 lg/mL streptomycin (Meiji Seika Pharma Co., Ltd.). The CSC complete medium (4Z0-500-R; DS Pharma Biomedical) is identical to a CSC medium (4Z3-500-R; DS Pharma Biomedical) containing 10% fetal bovine serum and culture boost growth factor. HRMECs were maintained by passage every 3 to 4 days, and the cells from passages 3 to 8 were used for the experiments.
8
PBMC Isolation and Allogenic Mixed Lymphocyte Reaction
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Peripheral blood mononuclear cells (PBMCs), isolated from heparinized blood by using a Ficoll-Hypaque density gradient (Separate-L, Muto Pure Chemicals Co. Ltd., Tokyo, Japan), were suspended in RPMI 1640 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated fetal bovine serum (Medical and Biological Laboratories Co., Ltd., Nagoya, Japan), 100 μg/mL streptomycin, and 100 U/mL penicillin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan). For the MLRs, 1.5 × 105 PBMCs were cultured with 5.0 × 104 allogenic PBMCs, which had been treated with irradiation of 40 Gy according to a previous method [16 (link)], and CB asbestos at 5 μg/mL. Following 2 days of the MLRs, IL-2 (Peprotech, Rocky Hill, NJ, USA) was added at 100 pg/mL or 1 ng/mL for 5 days. International Union Against Cancer (UICC) standard CB was kindly provided by the Department of Occupational Health at the National Institute for Occupational Health of South Africa [17 (link)]. All blood samples were taken from healthy volunteers who provided informed consent. The project was approved by the Institutional Ethics Committees of Kawasaki Medical School.
9
Expansion of Murine Embryonic Stem Cells
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Every time a new batch of EBRTcPTbx6 cells was thawed, 1.5 μg/mL of puromycin was added for one week before propagation. EBRTcPTbx6 cells were maintained with Glasgow minimum essential medium (GMEM, Sigma–Aldrich), 10% FBS (MP Biomedicals), 0.1 mM NEAA (Wako), 1 mM sodium pyruvate (Sigma–Aldrich), 0.1 mM 2-mercaptoethanol (Sigma–Aldrich), 10 ng/mL doxycycline (Wako), 100 U/mL penicillin (Meiji Seika Pharma) and 100 μg/mL streptomycin (Meiji Seika Pharma), supplemented with 1000 U/mL mouse LIF (ORF genetics), 3 μM CHIR-99021 (FOCUS Biomolecules), and 0.4 μM PD-0325901 (Adooq Bio Science). 1 × 105 EBRTcPTbx6 cells were inoculated to a 60 mm cell culture dish (TrueLine, Nippon Genetics) that had been plated with MMC (Wako)-treated SNL 76/7 feeder cells (obtained from Riken Bioresource Center) on the previous day. The cells were passaged every other day, and subjected to isolation from feeder cells using differential adhesive properties to a gelatin-coated cell culture dish before performing differentiation assays as described below.
10
Human Retinal Microvascular Endothelial Cell Culture
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Primary HRMECs (Cell Systems, Kirkland, WA, USA) were cultured as described in detail [52 (link)]. The HRMECs were maintained in complete classic medium supplemented with CultureBoost-R (Cell Systems, Kirkland, WA, USA) and 100 μg/mL of streptomycin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan) and 100 U/mL penicillin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan). Before the cell seeding, the culture dishes and well plates were precoated with an attachment factor (Cell systems, Kirkland, WA, USA). The cells were maintained at 37 °C in a humidified atmosphere with 5% CO2. Passage 6 to 10 were used for the experiments.
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