Streptomycin pen strep

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Streptomycin (pen/strep) is a broad-spectrum antibiotic commonly used in cell culture media. It is effective against a wide range of bacteria, including both Gram-positive and Gram-negative species.

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Streptomycin (24872 citations) Pen/Strep (3363 citations) Penicillin-Streptomycin (Pen-Strep) (236 citations) Streptomycin (Strep) (6 citations) Penicillin and streptomycin (pen/strep, (3 citations) Penicillin/Streptomycin (pen/strep) antibiotic/antimycotic mixture (3 citations) Streptomycin (pen/strep; p/s) (2 citations) Penicillin G and Streptomycin (pen/strep) (2 citations) Penicillin/streptomycin (Pen/Strep) broad-spectrum antibiotic cocktail (2 citations) Gibco® penicillin–streptomycin (pen–strep), (2 citations)

28 protocols using streptomycin pen strep

Cell Culture Media and Viability Assays

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All murine cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM – Life Technologies) supplemented with 10% Fetal Bovine Serum (FBS – Life Technologies) and 100 units per ml penicillin, and 100 μg/ml streptomycin (pen/strep) (Life Technologies). All human pancreatic cancer cells were maintained in RPMI 1640 (Life Technologies) supplemented with 10% FBS and 100 units per ml penicillin, and 100 μg/ml streptomycin (pen/strep) (Life Technologies).
Nutrient-defined media (Figure 7D) was made in DMEM (Life Technologies, A14430) and supplemented with 5% FBS, 4mM L-Glutamine (Life Technologies), 5mM D- glucose (Fisher chemicals) or 25mM D-galactose (BD) as indicated. Viable cell numbers were determined by cell counting (Figure 1C), using the CellTiter-Glo Luminescent Cell Viability assay according to manufacturer protocol (Promega - Figures 1D and7D), or using the CyQUANT Cell Proliferation Assay according to manufacturer protocol (ThermoFisher- Figure 1I). Each cell counting experiment was initially seeded at equal numbers in technical duplicate to triplicate per independent experiment.

Nagdas S., Kashatus J.A., Nascimento A., Hussain S.S., Trainor R.E., Pollock S.R., Adair S.J., Michaels A.D., Sesaki H., Stelow E.B., Bauer T.W, & Kashatus D.F. (2019). Drp1 Promotes KRas-Driven Metabolic Changes to Drive Pancreatic Tumor Growth. Cell reports, 28(7), 1845-1859.e5.

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Cell Culture Media and Viability Assays

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Analytical Standards of Bacopaside I and II

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The analytical standards bacopaside I (CAS No. 382148-47-2, 89.6% purity by HPLC, Lot no. 00002002-T17H) and bacopaside II (CAS No. 382146-66-9, 98% purity HPLC, Lot Number: 00002002-T17H), derived from the medicinal herb bacopa monnieri, were obtained from ChromaDex (Irvine, CA, USA), solubilised in methanol at 10 mM and 1.5 mM stock solutions, respectively, and stored at −20 °C. Cell lines MDA-MB-231, T47D, MCF7 and BT-474 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). Cells were maintained in complete medium, either in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Carlsbad, CA, USA) for MDA-MB-231, MCF7 and BT-474 or Roswell Park Memorial Institute (RPMI) 1640 medium (Life Technologies, Carlsbad, CA, USA) for T47D, containing 10% foetal bovine serum (FBS) (Corning, NY, USA), 200 U/mL of penicillin, 200 μg/mL of streptomycin (pen strep; Life Technologies, Carlsbad, CA, USA) and 2 mM L-alanyl-L-glutamine dipeptide (GlutaMAX Supplement; Life Technologies, Carlsbad, CA, USA). Cells were grown under standard culture conditions at 37° C with 5% CO₂ in air and used within 4 passages.

Palethorpe H.M., Smith E., Tomita Y., Nakhjavani M., Yool A.J., Price T.J., Young J.P., Townsend A.R, & Hardingham J.E. (2019). Bacopasides I and II Act in Synergy to Inhibit the Growth, Migration and Invasion of Breast Cancer Cell Lines. Molecules, 24(19), 3539.

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Routine Culture of SEES2 Embryonic Stem Cells

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SEES2 ESCs were routinely cultured on a feeder layer of freshly plated gamma-irradiated mouse embryonic fibroblasts (MEFs) isolated from ICR embryos at 12.5 d gestation and passaged 2 times before irradiation (30 Gy), in the ESC culture media^{16 (link),17 (link)}. The ESC media consisted of KNOCKOUT-Dulbecco’s modified Eagle’s medium (KO-DMEM) (Life Technologies, CA, USA; #10829-018) supplemented with 20% KNOCKOUT-Serum Replacement (KO-SR; #10828-028), 2 mM Glutamax-I (#35050-079), 0.1 mM non-essential amino acids (NEAA; #11140-076), 50 U/ml penicillin-50 μg/ml streptomycin (Pen-Strep) (#15070-063), 0.055 mM β-mercaptoethanol (#21985-023) and recombinant human full-length bFGF (#PHG0261) at 10 ng/ml (all reagents from Life Technologies).

Tsuneishi R., Saku N., Miyata S., Akiyama S., Javaregowda P.K., Ite K., Takashima N., Toyoda M., Kimura T., Kuroda M., Nakazawa A., Kasahara M., Nonaka H., Kamiya A., Kiyono T., Yamauchi J, & Umezawa A. (2021). Ammonia-based enrichment and long-term propagation of zone I hepatocyte-like cells. Scientific Reports, 11, 11381.

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Cytotoxicity Evaluation of Peptides

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HeLa cells were maintained in DMEM high glucose media with 4 mM L-glutamine (NyClone) and 100 U/mL penicillin, 100 μg/mL streptomycin (pen/strep) (Life Technologies), and 10% (v/v) inactivated fetal bovine serum (FBS) (NyClone). Cells were grown in 5% CO₂ at 37°C and detached from the culturing dish at 80% confluency using 0.025% trypsin-EDTA (NyClone) treatment. Effects of the peptide on cell viability were estimated by the MTS (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay according to the manufacturer’s protocol (Promega, USA) with minor modifications and as described [30 ]. Mouse breast cancer 4T1 cells were also cultivated for cytotoxicity evaluation of the peptides in a similar manner.

Mishra B., Wang X., Lushnikova T., Zhang Y., Golla R.M., Narayana J.L., Wang C., McGuire T.R, & Wang G. (2018). Antibacterial, Antifungal, Anticancer Activities and Structural Bioinformatics Analysis of Six Naturally Occurring Temporins. Peptides, 106, 9-20.

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Immunomodulatory Effects of MSCs and EVs

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MSCs or EVs were co-cultured in 96-well plates with splenocytes from BALB/c mice (0.3 M cells/well) and C57BL/6 mice (0.6 M cells/well) in 5% heat-inactivated FBS (Atlanta Biologicals) plus 100 units/mL penicillin and 100 mg/mL streptomycin (pen/strep; both from Life Technologies) in RPMI-1640 medium (ATCC). All mice were purchased from Jackson Laboratory. MSCs and splenocytes from BALB/c mice were pretreated with mitomycin (2.5 mg/mL for 2 hr at 37°C; Sigma-Aldrich) before co-culture. Two days or five days later, mouse cytokine expression was measured by real-time PCR assays or ELISAs according to the manufacturer's protocols.

Shigemoto-Kuroda T., Oh J.Y., Kim D.K., Jeong H.J., Park S.Y., Lee H.J., Park J.W., Kim T.W., An S.Y., Prockop D.J, & Lee R.H. (2017). MSC-derived Extracellular Vesicles Attenuate Immune Responses in Two Autoimmune Murine Models: Type 1 Diabetes and Uveoretinitis. Stem Cell Reports, 8(5), 1214-1225.

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Evaluation of Peptide Effects on HaCaT Cells

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Human keratinocytes from histologically normal skin (HaCaT cells; Fisher Cat # T0020001) were maintained in DMEM
High Glucose media with 4 mM L-Glutamine (NyClone) and 100 U/mL penicillin, 100 μg/mL streptomycin (pen/strep) (Life
Technologies), and 10% (v/v) inactivated fetal bovine serum (FBS) (NyClone). Cells were grown in 5% CO₂at 37°C and were detached from culturing dish at 80% confluency using 0.025% trypsin-EDTA (NyClone)
treatment. Peptide influence on the cell viability was estimated by using the MTS assay according to manufacturer’s
protocol (MTS, CellTiter96 AQ One Solution Cell Proliferation Assay, Promega).

Wang X., Mishra B., Lushnikova T., Narayana J.L, & Wang G. (2018). Amino Acid Composition Determines Peptide Activity Spectrum and Hot-Spot-Based Design of Merecidin. Advanced biosystems, 2(5), 1700259.

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Cell Culture and Drug Treatment

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HEK293T cells were kindly provided by Prof. Ron Kooijman (FARC, Vrije Universiteit Brussel) and were cultured in Dulbecco’s Modified Eagles Medium (DMEM) (Life Technologies, 31966-047) supplemented with 10% fetal bovine serum (FBS) (Perbio Science, SV30160.03) and 100 U/ml penicillin 100 μg/ml streptomycin (pen-strep) (Life Technologies, 15140-148). H1666 cells were purchased from the ATCC (CRL-5885) and cultured in F12-based (ATCC: 30-2006) ACL-4 medium supplemented with 10% FBS and pen-strep. Both cell lines were tested periodically for mycoplasma infection, and all tests were negative. Dabrafenib (Tafinlar) was provided by GlaxoSmithKline (UK). Trametinib (S2673) and AZ628 (S2746) were obtained from Selleckchem (USA).

Noeparast A., Giron P., De Brakeleer S., Eggermont C., De Ridder U., Teugels E, & De Grève J. (2018). Type II RAF inhibitor causes superior ERK pathway suppression compared to type I RAF inhibitor in cells expressing different BRAF mutant types recurrently found in lung cancer. Oncotarget, 9(22), 16110-16123.

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Cytotoxicity Evaluation of Peptides

Cited 1 time

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HeLa CCL-2 epithelial adenocarcinoma cells from American Type Culture Collection (ATCC) were maintained in DMEM high glucose media with 4 mM L-glutamine (NyClone) and 100 U/mL penicillin, 100 μg/mL streptomycin (pen/strep) (Life Technologies), and 10% (v/v) inactivated fetal bovine serum (FBS) (NyClone). Cells were grown in 5% CO₂ at 37°C and were detached from the culturing dish at 80% confluency using 0.025% trypsin-EDTA (NyClone) treatment. Peptide influence on the cell viability was estimated by using the MTS assay according to the manufacturer’s protocol (MTS, CellTiter96 AQ One Solution Cell Proliferation Assay, Promega) with minor modifications and as described elsewhere [20 (link)]. Human skin HaCaT (immortalized keratinocyte from AddexBio-T0020001) and kidney HEK293 cells were also cultivated for cytotoxicity evaluation of the peptides in a similar manner.

Mishra B., Lushnikova T., Golla R.M., Wang X, & Wang G. (2016). Design and surface immobilization of short anti-biofilm peptides. Acta biomaterialia, 49, 316-328.

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Cell Culture Maintenance Protocol

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A549 (ATCC, human, male), Vero (ATCC, primate, female), or HEp-2 cells (ATCC, human, female) were maintained in DMEM (Lonza) supplemented with 10% fetal bovine serum (FBS) (Hyclone) and 100 U/ml penicillin and 100 mg/ml streptomycin (pen-strep) (Life Technologies). All cells were cultured at 37°C and 5% CO₂. Cells were authenticated by ATCC and checked for mycoplasma contamination in our laboratory.

Blanchard E.L., Braun M.R., Lifland A.W., Ludeke B., Noton S.L., Vanover D., Zurla C., Fearns R, & Santangelo P.J. (2020). Polymerase-tagged respiratory syncytial virus reveals a dynamic rearrangement of the ribonucleocapsid complex during infection. PLoS Pathogens, 16(10), e1008987.

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