Vero (African green monkey kidney cells), HEp-2 (human laryngeal cancer/HELA contaminant), Panc 04.03 (human pancreatic cancer), and DU 145 (human prostatic cancer) were obtained from the American Type-Culture Collection (ATCC) (Rockville, MD, USA). L929-/-cGAS (in this paper called “L929KO”) and L929-/-cGAS reconstituted with human cGAS (called “L929R”) were a gift from Pinghui Feng; L929 cells are mouse subcutaneous fibroblasts. Veros, HEp-2, L929KO, and L929R, were maintained in Dulbecco’s Modified Eagle medium (Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin (Invitrogen, INC., Carlsbad, CA, USA). DU 145 cells were maintained on Minimum Essential medium supplemented with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin (Invitrogen, INC., Carlsbad, CA, USA). Panc 04.03 were maintained on RPMI-1640 complete medium (Gibco-BRL, Grand Island, NY, USA) supplemented with 20 U/mL recombinant human insulin, 15% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA), and 100 ug/mL Primocin (Invitrogen, INC., Carlsbad, CA, USA).
Primocin
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, China, Switzerland
Primocin is an antibiotic selection reagent used in cell culture applications. It is designed to inhibit the growth of bacteria, fungi, and mycoplasma, ensuring the purity of cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (31 mentions)
Glutamax, by Thermo Fisher Scientific (22 mentions)
Streptomycin, by Thermo Fisher Scientific (10 mentions)
Primocin, by InvivoGen (9 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions)
Nicotinamide, by Merck Group (9 mentions)
A83 01, by Bio-Techne (8 mentions)
L glutamine, by Thermo Fisher Scientific (8 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (6 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (6 mentions)
N acetyl l cysteine, by Merck Group (6 mentions)
Matrigel, by Corning (6 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Neurobasal medium, by Thermo Fisher Scientific (5 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (5 mentions)
Dmem f12, by Thermo Fisher Scientific (5 mentions)
Activin a, by Thermo Fisher Scientific (4 mentions)
Fgf10, by Thermo Fisher Scientific (4 mentions)
Insulin, by Merck Group (4 mentions)
Noggin, by Thermo Fisher Scientific (4 mentions)
106 protocols using primocin
1
Cell Line Maintenance for Cancer Research
2
Differentiation of hESCs into PGCLCs
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PGCLCs were induced from primed hESCs as described in Sasaki et al. (2015) (link), with some modifications (Chen et al., 2017b ). Day 7 hESCs were dissociated into single cells with 0.05% Trypsin-EDTA and plated onto Human Plasma Fibronectin (Invitrogen, 33016-015)-coated 12-well-plate at the density of 200,000 cells/well in 2 mL/well of iMeLC media, which is composed of 15% KSR, 1× NEAA, 0.1 mM 2-Mercaptoethanol, 1× PSG (Gibco, 10378-016), 1 mM sodium pyruvate (Gibco, 11360-070), 50 ng/mL Activin A (Peprotech, AF-120-14E), 3 μM CHIR99021 (Stemgent, 04-0004), 10 μM of ROCKi (Y27632, Stemgent, 04-0012-10), and 50 ng/mL primocin in Glasgow’s MEM (GMEM) (Gibco, 11710-035). iMeLCs were dissociated into single cells with 0.05% Trypsin-EDTA after 24 h of incubation and plated into ultra-low cell attachment U-bottom 96-well plates (Corning, 7007) at the density of 3000 cells/well in 200 μL/well of PGCLC media, which is composed of 15% KSR, 1× NEAA, 0.1 mM 2-Mercaptoethanol, 1× PSG (Gibco, 10378-016), 1 mM sodium pyruvate (Gibco, 11360-070), 10 ng/mL human LIF (Millipore, LIF1005), 200 ng/mL human BMP4 (R&D systems, 314-BP), 50 ng/mL human EGF (R&D systems, 236-EG) 10 μM of ROCKi (Y27632, Stemgent, 04-0012-10), and 50 ng/mL primocin in Glasgow’s MEM (GMEM) (Gibco, 11710-035).
3
Differentiation of hESCs into PGCLCs
4
Differentiating hESCs into hPGCLC
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Primed hESCs were differentiated into hPGCLC as described in Sasaki et al. (2015) (link) with some modifications. Day-7 hESCs were dissociated into single cells with 0.05% trypsin-EDTA (Gibco) and plated onto a human plasma fibronectin (Invitrogen)-coated 12-well-plate at 200,000 cells/well cell density in 2 mL/well of incipient mesoderm-like cells (iMeLCs) medium, which is composed of 15% knockout serum replacement (KSR), 1× non-essential amino acids (NEAA), 0.1 mM 2-mercaptoethanol, 1× penicillin-streptomycin-glutamine (Gibco), 1 mM sodium pyruvate (Gibco), 50 ng/mL Activin A (Peprotech), 3 μM CHIR99021 (Stemgent), 10 μM ROCKi (Y27632, Stemgent), and 50 ng/mL primocin in Glasgow's minimal essential medium (GMEM) (Gibco). Twenty-four hours later, iMeLCs were dissociated into single cells by 0.05% trypsin-EDTA, followed by plating onto ultra-low cell attachment U-bottom 96-well plates (Corning) at a density of 3,000 cells/well in 200 μL/well of hPGCLC medium, which is composed of 15% KSR, 1× NEAA, 0.1 mM 2-mercaptoethanol, 1× penicillin-streptomycin-glutamine (Gibco), 1 mM sodium pyruvate (Gibco), 10 ng/mL human leukemia inhibitory factor (Millipore), 200 ng/mL human BMP4 (R&D systems), 50 ng/mL human epidermal growth factor (R&D Systems), 10 μM of ROCKi (Y27632, Stemgent), and 50 ng/mL primocin in GMEM (Gibco). Day-4 hPGCLC aggregates were used for further analysis.
5
Hypoxic Stress Response of Hippocampal Neurons
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Primary neurons isolated from the hippocampus of postnatal 24 h NMRs were cultured in Dulbecco modified Eagle medium (DMEM) containing 10% heat‐inactivated foetal bovine serum (Life Technologies), 2 mM glucose, 0.0025% Glutamax, and 0.2 mg/ml primocin (Thermo Fisher Scientific). Primary neurons isolated from the hippocampus of postnatal 24 h mice were cultured in DMEM containing 10% heat‐inactivated foetal bovine serum (Life Technologies), 4.5 g/L glucose, 0.0025% Glutamax, and 0.2 mg/ml primocin (Thermo Fisher Scientific). Four hours later, the medium was replaced with neurobasal A medium supplemented with B27 (2%), 250 mM Glutamax, and penicillin/streptomycin (1%). NMR neurons were maintained at 33°C in 21% O2/5% CO2. Half of the medium was renewed every 48–72 h before further experiments. For hypoxic stress, hippocampal neurons from NMRs were maintained at 33°C (5% CO2, 8% O2), and hippocampal neurons from NMRs were maintained at 37°C (5% CO2, 8% O2), for the indicated periods of time in the absence or presence of 20 mM echinomycin (Sigma–Aldrich).
6
Characterization of Glioblastoma Cell Lines
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All cell lines were incubated at 37°C in an atmosphere containing 5% CO2. GBM12 and GBM14 (mesenchymal subtypte), GBM61 (proneural subtype), and GBM43 (classical subtype) cells were obtained from Dr. Jann Sarkaria (Mayo Clinic, Rochester, MN). U251 cells were obtained from Sigma. GL261 cells was obtained from Repository of Tumors and Tumor Cell Lines, NCI (Bethesda, MD). From ATCC, 293T (ATCC CRL-3216) was obtained. Cells were cultured in DMEM (Fisher Scientific, MT10013CV), 10% FBS (Gemini), and 100 µg/mL of primocin (Invivogen, ant-pm-1). For the treatment experiment, cells were cultured in DMEM containing 1.5% FBS. All the cells were performed with passages four to nine. Astrocyte was purchased from ScienceCell Research Laboratories and were cultured in the media in DMEM, 10% FBS, N-2 Supplement (Thermo Fisher, 17502048), and 100 µg/mL of primocin. NCH644 stem-like glioma cells (Cell Line Services, 820403) were cultured in StemPro NSC SFM (Thermo Fisher, A1050901) with 100 µg/mL primocin for maintenance and for drug treatment. The cell line repositories performed the authentications and Mycoplasma contamination was excluded by the original source.
7
Vero and SH-SY5Y Cell Culture Protocol
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African green monkey kidney cells (Vero) and SH-SY5Y were obtained from the American Tissue-Culture Collection (ATCC) (Rockville, MD, USA). Veros were maintained on Dulbecco’s Modified Eagle Medium (Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin© (Invitrogen, INC., Carlsbad, CA, USA). SH-SY5Y were maintained on Eagle’s Minimum Essential Medium (ATCC, Rockville, MD, USA) supplemented with 15% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin© (Invitrogen, INC., Carlsbad, CA, USA). The wildtype (WT) and Δ481N viruses were a gift from Prashant Desai (Johns Hopkins University, Baltimore, MD, USA). The mutants were made on a KOS background with a GFP on UL37. Viruses were grown and titrated on Vero cells [23 (link)].
8
Culturing and Maintaining Human Cell Lines
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hUCB-MSCs were provided by Kangstem Biotech GMP Center (South Korea). All experiments described below were conducted with hUCB-MSCs at passage 5. hUCB-MSCs were maintained with KSB-3 Basal medium (Kangstem Biotech, South Korea) supplemented with 10% fetal bovine serum (FBS; Gibco, USA) and 100 μg/ml primocin (In-vitrogen, USA). HaCaT (Addexbio, USA), human dermal fibroblast (HDF; Cell Applications, USA), and HEK293FT (ATCC, USA) cell lines were maintained with Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS and 100 μg/ml primocin (Invitrogen). Human LAD2 cells were cultured with StemPro-34 medium (Invitrogen) containing 100 ng/ml recombinant human stem cell factor/c-kit ligand (R&D Systems, USA) and 2 mM L-glutamine (Sigma, USA). All cells were maintained at 37℃ in a 5% CO2 incubator. Recombinant human EGF (rhEGF), human interferon (IFN)-γ, and human TNF-α were purchased from PeproTech (NJ, USA).
9
Intrahepatic Cholangiocarcinoma Cell Line Establishment
10
Hippocampal Neuron Primary Culture
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Hippocampi were dissected from both female and male postnatal day 1 (P1) pups from Actb-MBS-KI homozygous, and C57BL/6 wildtype (Koatech) mice. Hippocampi were digested with 0.25% Trypsin (Gibco) at 37 °C for 15 min. Trypsin was inactivated by incubating hippocampi in FBS containing plating medium (10% FBS (Gibco), 1x Glutamax (Gibco), 0.1 mg/ml Primocin (Invitrogen) in Neurobasal A medium (Gibco), followed by trituration and plating onto overnight poly-D-lysine-coated confocal dishes (SPL Life Science) in 5% CO2, 37 °C incubator. After 4 h of plating, 2 ml of B27 medium (Neurobasal A medium (Gibco) supplemented with 1xB27 (Gibco), 1x Glutamax, (Gibco) and 0.1 mg/ml Primocin (Invitrogen)) were added and neurons were grown in B27 medium until the experiment.
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