Streptomycin

Manufactured by Irvine Scientific

Sourced in United States

Streptomycin is a broad-spectrum antibiotic produced by the bacterium Streptomyces griseus. It is commonly used in microbiology and molecular biology laboratories as a selection agent for bacterial cultures.

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Lab products found in correlation

Penicillin, by Irvine Scientific (12 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (6 mentions) Hepes, by Thermo Fisher Scientific (4 mentions) 2 mercaptoethanol, by Merck Group (4 mentions) Fetal bovine serum (fbs), by Irvine Scientific (3 mentions) Fetal bovine serum (fbs), by Gemini Bio (3 mentions) Glutamine, by Irvine Scientific (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Brefeldin a, by BD (2 mentions) Rpmi 1640, by Irvine Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Irvine Scientific (2 mentions) Panc 1, by American Type Culture Collection (2 mentions) Mycoplasma stain assay kit, by Beyotime (2 mentions) Phenol red, by Irvine Scientific (2 mentions) F 12k medium, by American Type Culture Collection (2 mentions) L glutamine, by Irvine Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Um uc 3, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)

14 protocols using streptomycin

Bladder Cancer and Lymph Node Stromal Cells

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The human bladder cancer cell line UM-UC-3 was purchased from the American Type Culture Collection (ATCC, Manassas, VA) [34 (link)]. A LNSC line, HK [35 (link)], which resembles mesenteric LNSC functionally [16 (link), 24 (link), 36 (link)], was used. HK cells show the LNSC phenotype expressing CD320, CD44, α-SMA as well as CXCL12, IL-6, and IL-8 [16 (link)].
UM-UC-3 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) with 10% fetal bovine serum (FBS, Life Technologies, Grand Island, NY), 2 mM glutamine, 100 U/ml penicillin G, and 100 mg/ml streptomycin (Irvine Scientific, Santa Ana, CA) at 37° C in a 5% CO₂ humidified incubator. HK cells were maintained in RPMI media supplemented with 10% FBS, 2 mM glutamine, 100 U/ml penicillin G, and 100 mg/ml streptomycin [37 (link)]. HK cells can be isolated, grown, and expanded for up to 15 passages in vitro.

Gills J., Moret R., Zhang X., Nelson J., Maresh G., Hellmers L., Canter D., Hudson M., Halat S., Matrana M., Marino M.P., Reiser J., Shuh M., Laborde E., Latsis M., Talwar S., Bardot S, & Li L. (2018). A patient-derived orthotopic xenograft model enabling human high-grade urothelial cell carcinoma of the bladder tumor implantation, growth, angiogenesis, and metastasis. Oncotarget, 9(66), 32718-32729.

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Functional T Cell Subset Analysis

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Frozen PBMCs were thawed, washed twice in RPMI (Invitrogen) with 10% human AB serum, and rested at 37°C overnight. Cells were suspended in RPMI with 10% human AB serum, HEPES (Gibco BRL), glutamine, penicillin, streptomycin (Irvine Scientific), and 2‐mercaptoethanol (Sigma‐Aldrich) and cultured in 24‐well plates at a concentration of 1–3 × 10⁶ cells/mL with 50 ng/mL PMA (Sigma) and 1 mM ionomycin (Sigma). One microlitre of Brefeldin A (BD Bioscience) per millitre medium was added at the beginning of the culture. After 4 h cultured cells were washed twice. Th1/Tc1, Th2/Tc2, and Th17/Tc17 cells were identified by their intracellular co‐expression of IFN‐γ and the transcription factor T‐bet for Th1/Tc1,36 IL‐4 and GATA‐3 for Th2/Tc2,43 and IL‐17 and the transcription factor RORγt for Th17/Tc1744 (Table3C). The frequency of cells co‐expressing cytokines and transcription factors was measured by intracellular stain and flow cytometry. In addition, the relative frequency of CD4⁺ or CD8⁺ T cells that express IL‐2, IL‐4, IFN‐γ, TNF‐α, or IL‐17 was determined by intracellular stain. All cytokine‐positive T cell subsets were analysed in blinded fashion.

Narsale A., Moya R., Ma J., Anderson L.J., Wu D., Garcia J.M, & Davies J.D. (2019). Cancer‐driven changes link T cell frequency to muscle strength in people with cancer: a pilot study. Journal of Cachexia, Sarcopenia and Muscle, 10(4), 827-843.

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Culturing Human Pancreatic Cancer Cell Lines

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The PANC-1 and BxPC-3 human pancreatic cancer cell lines (American Type Culture Collection, Manassas, VA, USA) were respectively cultured in DMEM and RPMI 1640 medium supplemented with fetal bovine serum (10%), penicillin (100 U/ml), and streptomycin (100 mg/ml) (Irvine Scientific, Irvine, CA, USA). All cells were maintained at 37°C in humidified air with 5% CO_2. All other reagents were obtained from HyClone China Ltd., Beijing, China. Testing for mycoplasma contamination was done using a Mycoplasma Stain Assay Kit (Beyotime Institute of Biotechnology, Beijing, China). No cell cultures were contaminated with mycoplasma.

Li Y., Kong R., Chen H., Zhao Z., Li L., Li J., Hu J., Zhang G., Pan S., Wang Y., Wang G., Chen H, & Sun B. (2019). Overexpression of KLF5 is associated with poor survival and G1/S progression in pancreatic cancer. Aging (Albany NY), 11(14), 5035-5057.

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Isolation of Murine Reproductive Cells

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Reproductive tracts from GFP-CETN2-expressing males were collected intact into warm PBS after sacrificed by IACUC-approved protocols. After fat pad and mesenchymal tissues removal, the reproductive tracts were dissected into sections (testis, caput, upper corpus, middle upper corpus, mid-corpus, lower corpus, cauda, vas deferens) and placed individually into modified Human Tubal Fluid (HTF) medium without calcium or protein supplementation [mHTF-Hepes: 97.8 mM NaCl/4.69 mM KCl/0.20 mM MgSO₄▪7H₂O/0.37 mM KH₂PO₄/4.0 mM NaHCO₃/21.0 mM HEPES/2.78 mM glucose/21.4 mM sodium lactate/100 U/ml penicillin/100 μg/ml streptomycin/5 mg/liter phenol red; formulation modified from Irvine Scientific, Santa Ana, CA] kept at 37°. Testicular cells and epididymal sperm were extruded with fine forceps or a sterile 30-guage needle, with care taken to avoid sample mixing. Testicular cells and sperm suspensions were passed through sterile 40-μm nylon mesh filters (Fisher Scientific, Pittsburgh, PA) to remove large cellular debris, centrifuged at 400 × g for 10 minutes at room temperature, and pellets suspended in fresh warm mHTF-Hepes at 37 °C until use.

Simerly C., Castro C., Hartnett C., Lin C.C., Sukhwani M., Orwig K, & Schatten G. (2016). Post-Testicular Sperm Maturation: Centriole Pairs, Found in Upper Epididymis, are Destroyed Prior to Sperm’s Release at Ejaculation. Scientific Reports, 6, 31816.

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Aspergillus fumigatus Pathogenesis in A549 Cells

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Aspergillus fumigatus strains Af293 and CEA10 were grown on Sabouraud agar (Difco) at 37 °C for 5–7 days prior to use. The conidia were harvested by rinsing the plate with PBS containing 0.1 % Tween 80 (Sigma-Aldrich) and enumerated using a haemacytometer. For use in the experiments, the A549 type II pneumocyte cell line [American Type Culture Collection (ATCC)] was grown in F-12 K medium (ATCC) containing 10 % FBS (Gemini Bio-Products) and 1 % streptomycin and penicillin (Irvine Scientific) in 5 % CO₂ at 37 °C. The day before the experiments, approximately 7×10⁵ A549 cells in 2 ml of growth medium were cultured in each well of a six-well tissue culture plate. The next day, the A549 cells were gently rinsed twice with serum-free F-12 K medium. Next, 1×10⁶ conidia of each strain in 2 ml of F-12 K medium were added to individual wells of the six-well tissue culture plate. Control wells contained uninfected A549 cells and organisms incubated in F-12 K medium in the absence of A549 cells. After 6 and 16 h of incubation, RNA was extracted from the A549 cells and from A. fumigatus strains using the RiboPure Kit (Ambion, Life Technology) and purified using the RNA Clean and Concentrator Kit (Zymo Research).

Watkins T.N., Liu H., Chung M., Hazen T.H., Dunning Hotopp J.C., Filler S.G, & Bruno V.M. (2018). Comparative transcriptomics of Aspergillus fumigatus strains upon exposure to human airway epithelial cells. Microbial Genomics, 4(2), e000154.

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Intracellular Cytokine Profiling of T-Cell Subsets

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Either unsorted PBMC or sorted cell subsets, as indicated for each
experiment, were washed twice in RPMI (Invitrogen) with 10% human AB
serum and rested at 37°C overnight. Cells were resuspended in RPMI with
10% human AB serum, HEPES (Gibco BRL), glutamine, penicillin,
streptomycin (Irvine Scientific), and 2-mercaptoethanol (Sigma-Aldrich) and
cultured in 24 well plates at a concentration of 1–3 ×
10⁶ cell per ml with 50ng/ml PMA (Sigma) and 1μM
Ionomycin (Sigma). 1 μl of Brefeldin A (BD Bioscience) per ml medium was
added at the beginning of the culture. After 4 hours cultured cells were washed
twice. Th1 and Th2 cells were identified by their intracellular co-expression of
either IFN-γ and the transcription factor T-bet [25 (link)], or IL-4 and the transcription factor
GATA-3 [11 (link)],
respectively. Pro-inflammatory cytokines IL-2, TNF-α, and IL-17, as well
as the Th2-type cytokine IL-10 were also measured by intracellular stain and
Flow Cytometry.

Narsale A., Moya R, & Davies J.D. (2018). Human CD4+ CD25+ CD127hi cells and the Th1/Th2 phenotype. Clinical immunology (Orlando, Fla.), 188, 103-112.

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T Cell Cytokine Profiling Protocol

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Enriched or sorted CD4⁺ T cell subsets were incubated in triplicate at the concentration indicated for each experiment in 96-well plates and stimulated for 2 days with 1 μg/ml plate-bound anti-CD3 mAb (145-2C11), and 1 μg/ml soluble anti-CD28 (37.51) mAb (BD Biosciences). Cells were incubated in RPMI (Invitrogen) with 5% fetal bovine serum (Intergen), HEPES (Gibco BRL), glutamine, penicillin, streptomycin (Irvine Scientific), and 2-mercaptoethanol (Sigma-Aldrich). For cytokine analysis, 100 [.proportional]l of culture supernatant was collected from each well and concentrations of IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and TNF-α were determined by Flow Cytometry using the Th1/Th2/Th17 Cytometric Bead Array following the manufacturer's instructions (BD Biosciences). IL-22 and TGF-β were measured using ELISA kits according to the manufacturer's instructions (eBioscience). For intracellular cytokine staining, for the last four hours of culture, 1 [.proportional]l of BD GolgiPlug per ml medium was added to each culture and swirled gently to mix thoroughly. Cultured cells were washed twice and labeled with anti-CD4 mAb. Intracellular IL-4, IL-10, IL-17A, TNF-α, IL-2, IL-6, IFN-γ, and IL-22 content was determined by Flow Cytometry using cytokine-specific antibodies according to the manufacturer's instructions (BD Biosciences).

Zhao C., Marrero I., Narsale A., Moya R, & Davies J.D. (2015). CD4+ CD44v.low cells are unique peripheral precursors that are distinct from recent thymic emigrants and stem cell-like memory cells. Cellular immunology, 296(2), 106-114.

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Culturing Human Pancreatic Cancer Cell Lines

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The human pancreatic cancer cell lines PANC-1, BxPC-3, SW1990, and CFPAC-1 were obtained from the American Type Culture Collection (Rockville, USA) and were cultured in DMEM or RPMI 1640 medium supplemented with fetal bovine serum (10%), penicillin (100 U/mL), and streptomycin (100 mg/mL) (Irvine Scientific, Irvine, CA). All cells were maintained at 37°C in humidified air with 5% CO₂. All reagents were from HyClone China Ltd. (China). Mycoplasma contamination was tested using the Mycoplasma Stain Assay Kit (Beyotime Institute of Biotechnology, Beijing, China); none of the cell cultures were contaminated with mycoplasma.

Li Y., Wang Y., Kong R., Xue D., Pan S., Chen H, & Sun B. (2016). Dihydroartemisinin suppresses pancreatic cancer cells via a microRNA-mRNA regulatory network. Oncotarget, 7(38), 62460-62473.

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Preparation and Labeling of HTF Medium

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Human tubal fluid (HTF) medium (5.94 g/L NaCl, 0.35 g/L KCl, 0.05 g/L MgSO₄⋅7H₂O, 0.05 g/L KH₂ PO₄, 0.3 g/L CaCl₂ 2H₂O, 2.1 g/L NaHCO₃, 0.51 g/L D-glucose, 0.036 g/L Na pyruvate, 2.39 g/L Na lactate, 0.06 g/L penicillin, 0.05 g/L streptomycin, 0.01 g/L phenol red) was purchased from Irvine Scientific (Santa Ana, CA, USA). Pisum sativum lectin (PSL) labeled with fluorescein isothiocyanate (FITC-PSL), poly-l-lysine, bovine serum albumin (BSA), and 4′,6-diamidino-2-phenylindole were acquired from Sigma-Aldrich S.A. Ni-NTA-agarose was purchased from Qiagen (Tecnolab SA, Buenos Aires, Argentina); DL-dithiothreitol and isopropyl-β-D-1-thiogalactopyranoside (IPTG) were purchased from ICN (Eurolab SA, Buenos Aires, Argentina). Anti-GST antibody was from Calbiochem (MERCK, Buenos Aires, Argentina). Horseradish peroxidase-coupled anti-rabbit and Cy3-labeled goat anti-rabbit antibodies were from Jackson ImnunoResearch (Seroimmuno Diagnostics, Inc., Tucker, GA, USA). A23187 was from Alomone Labs. Ltd., Jerusalem, Israel. Pelco (Ted Pella Inc., CA, USA) supplied all electron microscopy supplies. All other chemical reagents were of analytical grade and were purchased from ICN or Sigma-Aldrich.

Berberian M.V., Pocognoni C.A, & Mayorga L.S. (2018). A TEM-traceable physiologically functional gold nanoprobe that permeates non-endocytic cells. International Journal of Nanomedicine, 13, 8075-8086.

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Rat Peritoneal Macrophage Isolation and Culture

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Rat peritoneal macrophages were harvested as previously described [23 (link)]. Macrophages were collected from 6-OHDA and control rats, following sacrifice, 4 and 8 weeks after 6-OHDA or saline injection. Cells were cultured as a monolayer in RPMI 1640 medium (Cellgro, Mediatech Inc., Herndon, VA), supplemented with 10 % fetal bovine serum (Gemini Bio-Products, Calabasas, CA), 2 mM L-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin (Irvine Scientific, Santa Ana, CA). Monolayers were washed 4 h after plating to remove non-adherent cells. Cultures were shown to be >98 % pure macrophages, as assessed by staining of non-specific esterase and macrophage-specific F4/80 mAb (data not shown).

Pellegrini C., Fornai M., Colucci R., Tirotta E., Blandini F., Levandis G., Cerri S., Segnani C., Ippolito C., Bernardini N., Cseri K., Blandizzi C., Haskó G, & Antonioli L. (2016). Alteration of colonic excitatory tachykininergic motility and enteric inflammation following dopaminergic nigrostriatal neurodegeneration. Journal of Neuroinflammation, 13, 146.

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