HEK293 cells (ATCC CRL-1573) were cultured in Eagle’s Minimum Essential Medium (ATCC, 30–2003), supplemented with 10% FBS (Corning, 35-011-CV) and 1% penicillin/streptomycin (Corning, 30-002-CI). AC16 human cardiomyocyte cell line (MilliporeSigma, SCC109) was cultured in DMEM/Hams F-12 50/50 Mix (Corning, 10-092-CV), supplemented with 12.5%FBS and 1% penicillin/streptomycin. HCF cells (Cell Applications Inc, 306V-05a) were cultured in HCF Growth Medium (Cell Applications Inc, 316–500).
Dmem hams f 12 50 50 mix
Manufactured by Corning
Sourced in United States
DMEM/Hams F-12 50/50 Mix is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types. It is a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture, providing a balanced formulation of nutrients, vitamins, and salts to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Corning (5 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Hek293 cell, by American Type Culture Collection (2 mentions)
Ac16 human cardiomyocyte cell line, by Merck Group (2 mentions)
Hcf growth medium, by Cell Applications (2 mentions)
Streptomycin, by GE Healthcare (2 mentions)
Geneticin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
L glutamine, by Merck Group (1 mentions)
Sv huc 1, by Corning (1 mentions)
Sv huc 1, by American Type Culture Collection (1 mentions)
Amphotericin b, by Corning (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Tempplate optical film, by USA Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Infinite m200, by Tecan (1 mentions)
Sodium bicarbonate solution, by Thermo Fisher Scientific (1 mentions)
12 protocols using dmem hams f 12 50 50 mix
1
Cell Culture Protocols for HEK293, AC16, and HCF
2
Immortalized Ovarian Cell Lines
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FTSECs expressing human telomerase reverse transcriptase and the SV40 large T and small T antigens were provided by Dr Ronny Drapkin (Dana-Farber Cancer Institute, Boston), and cultured in DMEM/Ham's F12 50/50 mix (Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (Corning). Additional introduction of GAB2 or a control vector into FTSECs were described previously.4 (link) FUOV1, NIH:OVCAR3 and IGROV1 cells were obtained and cultured as described.35 (link) These cell lines have been authenticated by sequenom genotyping assays for a panel of 48 single-nucleotide polymorphism loci with reference to the established fingerprint (http://www.broadinstitute.org/ccle ). No mycoplasma contamination was detected.
3
Cell Culture Protocols for Diverse Cell Lines
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HEK293 cells (ATCC CRL-1573) were cultured in Eagle’s Minimum Essential Medium (ATCC, 30–2003), supplemented with 10% FBS (Corning, 35-011-CV) and 1% penicillin/streptomycin (Corning, 30-002-CI). Lenti-X 293T and AAVpro 293T cell Lines were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with high glucose (4.5 g/L), 4 mM L-glutamine, and 3.7 g/L sodium bicarbonate (Sigma-Aldrich Co., No. D5796); supplemented with 10% fetal bovine serum and 1 mM sodium pyruvate. AC16 human cardiomyocyte cell line (MilliporeSigma, SCC109) was cultured in DMEM/Hams F-12 50/50 Mix (Corning, 10-092-CV), supplemented with 12.5%FBS and 1% penicillin/streptomycin. Human Cardiac Fibroblasts (HCF) (Cell Applications Inc, 306V-05a) were cultured in HCF Growth Medium (Cell Applications Inc, 316–500). Cell lines have been authenticated either by the vendor, or by our group using qPCR of cell-type specific gene markers. All cell lines were tested for mycoplasma contamination at the beginning of the projects and no contamination was observed.
4
Murine Triple Negative Breast Cancer Cell Lines
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Murine triple negative breast cancer cell lines TSA, luciferase-expressing 4T161 and EMT6 were kindly provided by Drs. Lizhong Wang and Runhua Liu, and Dr. Lalita Shevde-Samant, and Dr. Narendra Wajapeyee (all at University of Alabama at Birmingham), respectively. 4T1 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM)/Hams F-12 50/50 Mix (Corning, Cat# 10-090-CV) supplemented with 10% Fetal Bovine Serum (FBS) (MilliporeSigma, Cat# 12306C), and 10 units of Penicillin and 10 μg/mL Streptomycin (GE, Cat# SV30010), and 50 μg/mL Geneticin (ThermoFisher Scientific, Cat# 10131035). TSA and EMT6 cells were cultured in DMEM (MilliporeSigma, Cat# D6429) supplemented with 10% FBS, and 10 units of Penicillin and 10 μg/mL Streptomycin. All the cell lines used were confirmed pathogens free, including Mycoplasma, by Charles River Research Animal Diagnostic Services.
5
Culturing Bladder and Urothelial Cell Lines
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The human urinary bladder cancer cell line (T24) and human urothelium cell line (SV-HUC-1) were obtained from the American Type Culture Collection (ATCC, USA). The T24 cell line was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) Ham’s F-12 50/50 Mix (Corning, USA) and the SV-HUC-1 cell line in Kaighn’s Modification of Ham’s F-12 Medium (Corning, USA). Cells were kept at 37°C in a humidified atmosphere of 5% CO2. Both media were supplemented with 10% fetal bovine serum (FBS), 100 U/mL of penicillin-streptomycin, and 100 U/mL of amphotericin B (Corning, USA).
6
Murine Triple Negative Breast Cancer Cell Lines
7
Culturing MDA-MB-231 Breast Cancer Cells
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MDA-MB-231 human triple negative breast cancer cells (ATCC) were cultured in DMEM/Ham’s F12 50/50 mix (Corning) supplemented with 10% FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Corning). Cells were maintained at 37 °C and 5% CO2.
8
Circadian Transcription Factor Dynamics
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Real-time luciferase assays were performed in a 96-well plate format by reverse transfecting HEK293T (40,000 cells per well) with a total of 250 ng of DNA (10 ng of pGL3 Per1: Luc reporter, 30 ng CMV-CLOCK, 10 ng of CMV-Bmal1, and up to 200 ng of test plasmids) and 7.5 μl of TLT-1. The cells were seeded out in phenol red–free DMEM/Ham’s F-12 50/50 mix (Corning Cat# 16-405-CV) supplemented with 10% FBS, 1% Antibiotic-Antimycotic (Life Technologies), 25 mM HEPES, and 125 μM of D-Luciferin. The plate was sealed tight with TempPlate Optical film (USA Scientific). The plate was immediately transferred to the Tecan Infinite M200 maintained at 37 °C, and luminescence was measured in kinetic mode (every 20 minutes) for at least 72 hours. To determine the relative expression of flag components, lysates were prepared at the time corresponding to the peak of CLOCK–Bmal1 activity and analyzed by western blots.
9
Infection of Human RPE Cells
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Human RPE cells were grown in DMEM/F12 (Corning DMEM Hams F-12 50/50 Mix), supplemented with 10% of fetal bovine serum and 3% of sodium bicarbonate solution (Gibco) at 37°C with 5% CO2. Prior to infection, 2.5 × 105 cells⋅ ml–1 of RPE were seeded in 24-well plates to reach ∼80% confluence (microscopically verified) in 24 h. Media were gently aspirated from the top of the cells, and each well was washed using antibiotic-free media. Overnight LB liquid culture-grown bacteria were diluted in DMEM/F12 to achieve the multiplicity of infection (MOI) of 50:1. Infected cells were then incubated at 37°C for 24 h. Bacterial numbers were enumerated by plating serial dilutions onto LB agar and counting colonies at 0 (to verify MOI) and 24 h post-infection.
10
Isolation and Culture of Mouse Embryonic Fibroblasts
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