Penicillin streptomycin

Manufactured by Promega

Sourced in United States

Penicillin/streptomycin is a solution of the antibiotics penicillin and streptomycin. It is commonly used in cell culture applications to prevent bacterial contamination.

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Lab products found in correlation

Fugene hd transfection reagent, by Promega (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) B27 supplement, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) Glutamax supplement, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) Andlipofectamine 3000 reagent, by Thermo Fisher Scientific (2 mentions) Inopti mem medium, by Thermo Fisher Scientific (2 mentions) Papain, by Worthington (2 mentions) Puromycin, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Mtt assay, by Promega (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Thermolysin, by Promega (1 mentions) Dmem glutamax, by Thermo Fisher Scientific (1 mentions) Mouse 2.5 s ngf, by Promega (1 mentions) Opti mem medium, by Thermo Fisher Scientific (1 mentions) Dual glo luciferase assay system, by Promega (1 mentions)

Spelling variants of this product

Streptomycin (12 citations) Penicillin (11 citations)

12 protocols using penicillin streptomycin

Preparation and Transfection of Cortical Neuron Cultures

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Cortical neuron cultures were prepared from E16.5 mouse embryos.
Neocortices were dissociated by addition of papain (Worthington Biochemical,
LK003176) for 30 min at 37 °C. 250,000 cells/wells were plated in 12-well
plates and they were maintained in Neurobasal Medium (Gibco, 21103-049)
containing 2% B27 supplement (Gibco, 17504-044), 1% GlutaMAX supplement (Gibco,
35050-038), and 1% penicillin/streptomycin (Sigma, P4333). At DIV7, cortical
cultures were transfected with 400 ng/well of splicing reporters and
Lipofectamine 3000 reagent (ThermoFisher Scientific, L3000008) diluted in
opti-MEM medium (Gibco, 31985-062) using a 1:1.5 DNA-Lipofectamine ratio.
20,000 Neuroblastoma 2a (Neuro2a) or HEK293T cells (obtained from ATCC)
were plated in 96 well plates and were kept in DMEM (Sigma, D5796) supplemented
with 10% FBS (Gibco, 10270106) and 1% penicillin/streptomycin at 37°C.
After 24h, cells were transfected using FuGENE HD Transfection reagent (Promega,
E2691) with 50 ng of splicing reporter DNA alone or in combination with 50 ng of
splicing factor DNA.

Furlanis E., Traunmüller L., Fucile G, & Scheiffele P. (2019). Landscape of ribosome-engaged transcript isoforms reveals extensive neuronal cell class-specific alternative splicing programs. Nature neuroscience, 22(10), 1709-1717.

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Preparation and Transfection of Cortical Neuron Cultures

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Cytotoxicity Evaluation of MSNs and ICG/MSNs-RGD

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The cellular cytotoxicity of the MSNs and ICG/MSNs-RGD was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Promega, China). GFP-transfected luciferase-expressing human hepatocellular carcinoma cells (Hep-G2-GFP-fLuc cells), which expressed αvβ3 receptors and MDA-MB-231-fLuc human mammary cancer cells were purchased from the Academy of Military Medical Sciences (China) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, China) supplemented with 10% FBS and 1% penicillin/streptomycin (Promega) at 37 °C in an atmosphere containing 5% CO₂. Cells were seeded into 96-well microtiter plates at 10,000 cells per well. After incubation for 24 h, the cells were washed with phosphate-buffered saline (PBS), and the medium was replaced with fresh medium containing various concentrations of the MSNs or ICG/MSNs-RGD (30, 50, 100, 150, 200, 250, 300, 350, and 400 μg/mL). An equivalent volume of fresh medium (200 μL/well) was added to the control wells. After 24 h incubation, the treated cells were washed with PBS, and then fresh medium (100 μL) and MTT solution (10 μL, 5 mg/mL in PBS) were added to each well and cultured for another 4 h. The old medium was removed, and dimethyl sulfoxide (DMSO; 120 μL; China) was added to each well. The absorbance at 490 nm was read using a microplate reader to calculate relative cell viabilities.

Zeng C., Shang W., Wang K., Chi C., Jia X., Fang C., Yang D., Ye J., Fang C, & Tian J. (2016). Intraoperative Identification of Liver Cancer Microfoci Using a Targeted Near-Infrared Fluorescent Probe for Imaging-Guided Surgery. Scientific Reports, 6, 21959.

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Isolation of Hepatocytes and Non-Parenchymal Cells

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A cohort of 11-week-old ND-fed male Napepld^∆Hep and WT mice were anesthetized with Ketamine-Xylazine (Nimatek, Eurovet Animal Health BV, Bladel, Netherlands-Rompun, Bayer Healthcare, Loos, France) solution. The mice were surgically opened and the liver was infused with a PBS solution containing EGTA and HEPES buffer injected in the portal vein via a 26 g needle at a rate of 9 mL/min for 10 min. Warm (37 °C) digestion medium including DMEM/F12, no glutamine (Thermo Fisher, Waltham, MA, USA), Penicillin-Streptomycin, HEPES buffer, thermolysin (Promega, Madison, WI, USA), collagenase G (Abiel, Palermo, Italy) and collagenase H (Abiel, Palermo, Italy) was then added for 7 min. The liver was then removed and incubated with the medium at 37 °C for 15 min for further digestion. The obtained solution was then filtered (70 µm) and centrifuged twice (4 °C, 50 g, 2 min) to allow the separation between hepatocytes and non-parenchymal cells (NPC). The resulting pellet contained the hepatocytes whereas NPC were enriched in the supernatant. After isolating the two fractions in different tubes, they were centrifuged (4 °C, 10,000× g, 10 min) and pellets were stored in −80 °C prior TriPure RNA extraction.

Lefort C., Roumain M., Van Hul M., Rastelli M., Manco R., Leclercq I., Delzenne N.M., Marzo V.D., Flamand N., Luquet S., Silvestri C., Muccioli G.G, & Cani P.D. (2020). Hepatic NAPE-PLD Is a Key Regulator of Liver Lipid Metabolism. Cells, 9(5), 1247.

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Isolation and Culture of Rat Dorsal Root Ganglia

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DRG were harvested from neonatal Wistar rats (3–5 days old). Ganglia were digested with 0.25% (w/v) collagenase (type IA) in DMEM-glutamax (Invitrogen) with 1% penicillin-streptomycin (5000 U/mL, Invitrogen) for 1 h (37 °C, 5% CO₂). After digestion, rat DRG was mechanically dissociated using a glass Pasteur pipette. Single cell suspension was passed through a 100 μm cell strainer, and washed with DMEM glutamax plus 10% fetal bovine serum (FBS; Invitrogen) and 1% penicillin-streptomycin. Cells were seeded at the 30 μL of medium containing cells on MEA chambers previously coated with poly-L-lysine (8.33 μg/mL) and laminin (5 μg/mL). After 2 h, medium was replaced with DMEM glutamax, 10% FBS and 1% penicillin-streptomycin, supplemented with mouse 2.5s NGF 50 ng/mL (Promega), and 1.25 μg/mL cytosine arabinoside when required (37°C, 5% CO₂). All experiments were made 48 h after cell seeding.

González-Gil I., Zian D., Vázquez-Villa H., Hernández-Torres G., Martínez R.F., Khiar-Fernández N., Rivera R., Kihara Y., Devesa I., Mathivanan S., del Valle C.R., Zambrana-Infantes E., Puigdomenech M., Cincilla G., Sanchez-Martinez M., de Fonseca F.R., Ferrer-Montiel A.V., Chun J., López-Vales R., López-Rodríguez M.L, & Ortega-Gutiérrez S. (2019). A novel agonist of the type 1 lysophosphatidic acid receptor (LPA1), UCM-05194, shows efficacy in neuropathic pain amelioration. Journal of medicinal chemistry, 63(5), 2372-2390.

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Transcriptional Regulation Assay Protocol

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Trypsin-EDTA was purchased from Hyclone. opti-MEM medium was from Gibco-BRL (Invitrogen Life Technologies, Carlsbad, USA). Puromycin (Merck Calbiochem, Darmstadt, Germany), polybrene (Sigma-Aldrich, St. Louis, MO, USA), penicillin-streptomycin, Dual-Glo™ Luciferase Assay system (Promega, Madison, WI, USA), Lipofectamine 2000 with Plus (Life Technologies, Carlsbad, CA, USA) and recombinant human TGF-β1 (carrier-free; 580706; Biolegend, San Diego, CA, USA) were used. Antibodies used in the present study are listed in Table I.

WU Y.H., AI X., LIU F.Y., LIANG H.F., ZHANG B.X, & CHEN X.P. (2015). c-Jun N-terminal kinase inhibitor favors transforming growth factor-β to antagonize hepatitis B virus X protein-induced cell growth promotion in hepatocellular carcinoma. Molecular Medicine Reports, 13(2), 1345-1352.

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Immortalized Myoblast Culture Protocol

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RD-DUX4i cells were grown in DMEM (Gibco) supplemented with 10% FBS (Hyclone), 1% penicillin/streptomycin (Gibco) and 2.0 μg/ml puromycin (Sigma). The DUX4 transgene was induced with 1.0 μg/ml of doxycycline hyclate (Sigma). Unaffected (MB135) and FSHD (MB200) immortalized human myoblasts were cultured in F10 medium (Gibco/Life Technologies) supplemented with 20% FBS and 1% penicillin/streptomycin as well as 10 ng/ml recombinant human FGF (Promega) and 1 μM dexamethasone (Sigma). Human myoblasts were differentiated by culturing in knockout serum replacement medium (Gibco/Life Technologies).

Shadle S.C., Zhong J.W., Campbell A.E., Conerly M.L., Jagannathan S., Wong C.J., Morello T.D., van der Maarel S.M, & Tapscott S.J. (2017). DUX4-induced dsRNA and MYC mRNA stabilization activate apoptotic pathways in human cell models of facioscapulohumeral dystrophy. PLoS Genetics, 13(3), e1006658.

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LDLR and PCSK9 NanoBiT Assay

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The full-length coding sequence for LDLR was cloned into the pNB3K NanoBiT cloning vector (Promega) to yield the fusion protein LgBiT-GSSGGGGSGGGGSSG-AIA-LDLR. The native LDLR signal sequence was removed and replaced with the interleukin-6 signal sequence (amino acid sequence: MNSFSTSAFGPVAFSLGLLLVLPAAFPAP). The full-length coding sequence for PCSK9 was cloned into the pNB2K NanoBiT cloning vector (Promega) to yield the fusion protein PCSK9-VSQ-GSSGGGGSGGGGSSG-SmBiT-Histag. HEK293 transfection was carried out using FuGENE® HD Transfection Reagent (Promega) at a lipid:DNA ratio of 3:1. After 48 h, media with transfection reagent were removed and replaced with DMEM media (Thermo Fisher Scientific) containing 10% FBS, penicillin/streptomycin, and 400 µg/ml G418 (Promega). A pool of cells stably transfected with LgBiT-LDLR were selected that gave a high signal in the PCSK9-LDLR binding assay. The LgBiT-LDLR cells were subjected to fluorescence-activated cell sorting analysis to interrogate the integrity of the receptor. The LgBiT-LDLR was labeled with anti-LDLR antibody, which suggests that the receptor localized to the cell membrane and displayed as expected. A pool of cells stably transfected with PCSK9-SmBiT were selected and maintained. Stability of both stable cell pools was confirmed to 20 passages.

Duellman S.J., Machleidt T., Cali J.J, & Vidugiriene J. (2017). Cell-based, bioluminescent assay for monitoring the interaction between PCSK9 and the LDL receptor. Journal of Lipid Research, 58(8), 1722-1729.

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Expansion and Maintenance of Mouse Retinal Progenitor Cells

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Mouse RPCs were isolated from dissociated retinas of post-natal (P4) C57BL/6J mice, homozygous for expression of enhanced green fluorescent protein under control of the beta actin promoter (EGFP+/+) (gift from Dr Young, Harvard University^{52 (link)}). RPCs were expanded and maintained at 37°C in a 5% CO₂ incubator in neurobasal complete medium (Invitrogen-Gibco, Rockville, MD) containing 2 mM l-glutamine, 100 mg/mL penicillin-streptomycin, B27 and N2 neural supplements, and 20 ng/mL epidermal growth factor (Promega, Madison, WI) as described previously by our group.^{11 (link),27 (link)} For testing, cells were detached with trypsin-EDTA, re-suspected in culture medium, counted, and seeded as described below.

Thakur A., Mishra S., Pena J., Zhou J., Redenti S., Majeska R, & Vazquez M. (2018). Collective adhesion and displacement of retinal progenitor cells upon extracellular matrix substrates of transplantable biomaterials. Journal of Tissue Engineering, 9, 2041731417751286.

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Transfecting 293T17 and Ba/F3 Cells

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293T17 cells (ATCC) were maintained in DMEM (Invitrogen) supplemented with 10% FBS (Atlanta Biologicals), L-glutamine, penicillin/streptomycin (Invitrogen), and amphotericin B (HyClone). Ba/F3 cells were maintained in RPMI 1640 supplemented with 10% FBS, 15% WEHI-conditioned media (IL3 source), L-glutamine, penicillin/streptomycin, and amphotericin B. FuGENE 6 (Promega) was used for the transfection of 293T17 cells. Production of murine retrovirus was achieved by co-transfection of 293T17’s with CSF3R-MSCF-IRES-GFP constructs with pEcopac Helper and harvesting supernatants 48 hours post-transfection.

Maxson J.E., Luty S.B., MacManiman J.D., Paik J.C., Gotlib J., Greenberg P., Bahamadi S., Savage S.L., Abel M.L., Eide C.A., Loriaux M.M., Stevens E.A, & Tyner J.W. (2015). The Colony Stimulating Factor 3 Receptor T640N mutation is oncogenic, sensitive to JAK inhibition, and mimics T618I. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(3), 757-764.

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