Streptomycin

Manufactured by Corning

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Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It is produced by the actinobacterium Streptomyces griseus and is effective against a wide range of Gram-positive and Gram-negative bacteria. Streptomycin functions by inhibiting protein synthesis in bacterial cells, making it a useful tool for researchers and scientists working in microbiology and related fields.

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Lab products found in correlation

Penicillin, by Corning (787 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (177 mentions) L glutamine, by Corning (152 mentions) Fetal bovine serum (fbs), by Corning (119 mentions) Dulbecco modified eagle medium (dmem), by Corning (89 mentions) Rpmi 1640, by Corning (64 mentions) Fetal bovine serum (fbs), by Merck Group (49 mentions) Streptomycin, by Thermo Fisher Scientific (47 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (46 mentions) Amphotericin b, by Corning (44 mentions) Rpmi 1640 medium, by Corning (43 mentions) Penicillin, by Thermo Fisher Scientific (42 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (41 mentions) Glutamax, by Thermo Fisher Scientific (40 mentions) Sodium pyruvate, by Corning (38 mentions) Penicillin streptomycin, by Corning (33 mentions) Glutamine, by Corning (30 mentions) Hepes, by Corning (30 mentions) Penicillin g, by Corning (28 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (28 mentions)

865 protocols using streptomycin

Cell Culture Conditions for Infection and EV Collection

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Cells. Spinner-adapted L cells were grown in Joklik’s minimum essential medium (JMEM; U.S. Biological) supplemented to contain 5% fetal bovine serum (FBS; Gibco), 2 mM L-glutamine (Corning), 100 U/ml penicillin (Corning), 100 mg/ml streptomycin (Corning), and 25 ng/ml amphotericin B (Corning). During infection and EV collection, L cells were cultured in serum-free JMEM. Caco-2 cells were maintained in minimum essential medium (MEM; Corning) supplemented to contain 20% FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, 100 U/ml non-essential amino acids (Corning), 100 U/ml HEPES buffer (Corning), 100 U/ml sodium pyruvate (Corning), and 25 ng/ml amphotericin B. During infection and EV collection, Caco-2 cells were cultured in serum-free MEM and kept in a non-polarized, non-differentiated state through maintenance splitting and seeding at sub-confluent levels. Baby hamster kidney cells expressing T7 RNA polymerase controlled by a cytomegalovirus promoter (BHK-T7) were maintained in Dulbecco’s minimum essential medium (DMEM; Corning) supplemented to contain 5% FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, and 1 mg/ml Geneticin, which was added every other passage. All cells were maintained at 37°C with 5% CO₂.

Smith S.C., Krystofiak E, & Ogden K.M. (2024). Mammalian orthoreovirus can exit cells in extracellular vesicles. PLOS Pathogens, 20(1), e1011637.

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Cell Line Maintenance Protocol

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Human EBV transformed lymphoblastoid cell lines, GM06990 (control) and GM03200 (Fragile X), and fibroblast cell lines, GM00357 (control) and GM05848 (Fragile X), were purchased from Corielle institute. Lymphoblastoids were grown in RPMI1640 (Corning), supplemented with GlutaMAX (GIBCO), 15% heat-inactivated FBS (Fetal Bovine Serum, Benchmark), 100 IU/mL penicillin and 100 μg/mL streptomycin (Corning) at 37°C with 5% CO₂. Fibroblast cells were cultured in MEM culture media with 15% FBS (Corning), 1X GlutaMAX, 100 IU/mL penicillin and 100 μg/mL streptomycin. Cell lines were verified for presence or absence of FMRP and expansion of the FMR1 5′UTR using western blot and Southern blot respectively. See Figures 1A and S1A for results. Phoenix-AMPHO producer cells (ATCC) were grown in DMEM medium (GIBCO) supplemented with 10% FBS, 1X GlutaMAX, 100 IU/mL penicillin and 100 μg/mL streptomycin, 1mM sodium pyruvate (Corning), 10mM HEPES buffer (Corning) and 1X MEM non-essential amino acids (Corning).

Chakraborty A., Jenjaroenpun P., Li J., El Hilali S., McCulley A., Haarer B., Hoffman E.A., Belak A., Thorland A., Hehnly H., Schildkraut C., Chen C.L., Kuznetsov V.A, & Feng W. (2020). Replication Stress Induces Global Chromosome Breakage in the Fragile X Genome. Cell reports, 32(12), 108179.

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Culturing Oral Keratinocytes and KSHV Production

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The oral mucosal keratinocyte cell line OKF6/Tert-1 (Dickson et al., 2000 (link)) was cultured in keratinocyte serum-free media (K-SFM) (Gibco) supplemented with 2 mM L-glutamine (Corning), 1 IU/ml penicillin (Corning), 100 μg/ml streptomycin (Corning), 75 μg/ml bovine pituitary extract (Gibco), 0.3 mM CaCl₂ (Sigma-Aldrich), and 0.6 ng/ml epidermal growth factor (EGF) (Gibco). To produce KSHV, we used the iSLK-BAC16 cell line carrying the recombinant KSHV clone BAC16 (Brulois et al., 2012 (link)). The iSLK cell line is engineered to express the KSHV lytic switch gene RTA under doxycycline control (Myoung and Ganem, 2011b (link)). The resulting iSLK cell line was then further engineered to carry the KSHV clone BAC16 expressing GFP under the constitutive EF-1α human promoter (Brulois et al., 2012 (link)). The iSLK-BAC16 cells were cultured in Dulbecco’s modified Eagle’s media (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco), 1 IU/ml penicillin (Corning), 100 μg/ml streptomycin (Corning), and 1 mg/ml hygromycin B (Sigma-Aldrich). hygromycin B is used for the selection of cells with BAC16. HEK293 cells were grown in DMEM (Gibco) supplemented with 10% FBS (Gibco), 1 IU/ml penicillin (Corning), and 100 μg/ml streptomycin (Corning).

Brice D.C., Toth Z, & Diamond G. (2018). LL-37 Disrupts the Kaposi’s Sarcoma-Associated Herpesvirus Envelope and Inhibits Infection in Oral Epithelial Cells. Antiviral research, 158, 25-33.

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Cell Culture Fixation and Glycan Labeling

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C3H10T1/2 cells (ATCC # CCL-226) were grown in MEM NEAA Earle’s Salts (Irvine Scientific, #9130), supplemented with 10% fetal bovine serum (Corning, #35-015-CV), 2 mM l-glutamine, 100 units/mL penicillin and 0.1 mg/mL streptomycin (Sigma, #G6784). HUVEC (Clonetics/Lonza, #C2517A) were grown in EGM-2 Bullet Kit (Clonetics/Lonza, #CC-3162). Jurkat (John Blenis-Harvard Medical School, 240 Longwood Ave, Boston, MA 02,115) were grown in IMDM (ThermoFisher, #12,440,061), supplemented with 10% fetal bovine serum (Corning, #35-015-CV), 100 units/mL penicillin and 0.1 mg/mL streptomycin (Sigma, #G6784). CHO-K1 Wild Type and CHO-K1 pgsA-745 (ATCC # CRL-2242) were grown in IMDM (ThermoFisher, #12,440,061), supplemented with 5% fetal bovine serum (Corning, #35-015-CV), 100 units/mL penicillin and 0.1 mg/mL streptomycin (Sigma, #G6784). Upon confluence, cells were trypsinized and plated in a 24-well cell culture plate and grown to desired confluence. The cells were rinsed with sterile phosphate buffered saline (PBS) and fixed in 4% paraformaldehyde for 30 min at room temperature followed by washing five times with sterile PBS. After washing, the plate was stored in 1mL sterile PBS at 4°C until ready for glycan labeling.

Wu Z.L., Person A.D., Anderson M., Burroughs B., Tatge T., Khatri K., Zou Y., Wang L., Geders T., Zaia J, & Sackstein R. (2017). Imaging specific cellular glycan structures using glycosyltransferases via click chemistry. Glycobiology, 28(2), 69-79.

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Culturing Cell Lines for Extracellular Vesicle Isolation

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Spinner-adapted L cells were grown in Joklik’s minimum essential medium (JMEM; U.S. Biological) supplemented to contain 5% fetal bovine serum (FBS; Gibco), 2 mM L-glutamine (Corning), 100 U/ml penicillin (Corning), 100 mg/ml streptomycin (Corning), and 25 ng/ml amphotericin B (Corning). During infection and EV collection, L cells were cultured in serum-free JMEM. Caco-2 cells were maintained in minimum essential medium (MEM; Corning) supplemented to contain 20% FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, 100 U/ml non-essential amino acids (Corning), 100 U/ml HEPES buffer (Corning), 100 U/ml sodium pyruvate (Corning), and 25 ng/ml amphotericin B. During infection and EV collection, Caco-2 cells were cultured in serum-free MEM and kept in a non-polarized, non-differentiated state through maintenance splitting and seeding at sub-confluent levels. Baby hamster kidney cells expressing T7 RNA polymerase controlled by a cytomegalovirus promoter (BHK-T7) were maintained in Dulbecco’s minimum essential medium (DMEM; Corning) supplemented to contain 5% FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, and 1 mg/ml Geneticin, which was added every other passage. All cells were maintained at 37°C with 5% CO₂.

Smith S.C., Krystofiak E, & Ogden K.M. (2023). Mammalian orthoreovirus can exit cells in extracellular vesicles. bioRxiv.

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Generation and Maintenance of Cell Lines

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H3122 cells were obtained from the National Cancer Institute. Generation of ceritinib-resistant H3122 cells is described in the Supplemental Experimental Procedures. H2228, HCC827, and HCC4006 were purchased from the American Type Culture Collection (ATCC). PC-9 cells were obtained from Public Health England, and KELLY neuroblastoma cells were obtained from Sigma-Aldrich. Cells were maintained in RPMI-1640 (Cellgro) with 10% fetal bovine serum (Gemini Bioproducts) and penicillin (100 units/mL) / streptomycin (100 µg/mL; Cellgro). MGH006 cells have been previously reported and were maintained in DMEM (Cellgro) with 10% fetal bovine serum, penicillin, and streptomycin (Sequist et al., 2010 (link)). All cell lines were tested to confirm the absence of mycoplasma contamination. Crizotinib, TAE684, ceritinib, erlotinib, lapatinib, and sotrastaurin were purchased from Selleck Chemicals. Blasticidin was obtained from Life Technologies. Phorbol 12-myristate 13-acetate (PMA) was purchased from Santa Cruz Biotechnology.

Wilson F.H., Johannessen C.M., Piccioni F., Tamayo P., Kim J.W., Van Allen E.M., Corsello S.M., Capelletti M., Calles A., Butaney M., Sharifnia T., Gabriel S.B., Mesirov J.P., Hahn W.C., Engelman J.A., Meyerson M., Root D.E., Jänne P.A, & Garraway L.A. (2015). A Functional Landscape of Resistance to ALK Inhibition in Lung Cancer. Cancer cell, 27(3), 397-408.

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Doxycycline-Inducible Diffuse Large B-Cell Lymphoma Cell Lines

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Doxycycline-inducible human diffuse large B-cell lymphoma cell lines (HBL1, OCI-Ly10, and TMD8) that express the bacterial tetracycline repressor were engineered as described previously (27 (link)). Doxycycline (20 ng/ml) was used for inducing the expression of genes of interest. The cell lines were grown in RPMI 1640 media (Hyclone) supplemented with 20% FBS (fetal bovine serum, Atlanta Biologicals), 100 U/ml penicillin, 100 μg/ml streptomycin (Corning Cellgro), 2 mM GlutaGROTM (Corning Cellgro), 1×MEM-NEAA (Quanlity Biological, Inc.), and 1 mM Sodium Pyruvate Solution (Hyclone). All cultures were routinely tested for mycoplasma contamination. Human embryonic kidney cell line 293T was cultured in DMEM (Dulbecco’s modified Eagle’s medium, Hyclone) with 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin (Corning Cellgro). All cell lines were cultured at 37°C in a 5% CO₂ atmosphere.

Zhu F., Hwang B., Miyamoto S, & Rui L. (2016). Nuclear Import of JAK1 is Mediated by a Classical NLS and is Required for Survival of Diffuse Large B-cell Lymphoma. Molecular cancer research : MCR, 15(3), 348-357.

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Culturing Leukemia and Mammary Carcinoma Cells

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The human chronic myelogenous leukemia (CML) cell line K562 and human acute myeloid leukemia (AML) cell line Molm14 were cultured in RPMI 1640 (Corning, United States) supplemented with 10% fetal bovine serum (FBS, GIBCO, United States), 100 U/ml penicillin and 100 µg/ml streptomycin (Cellgro, Corning, United States) at 37°C with 5% CO₂. Murine 4T1 mammary carcinoma cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Corning, United States) supplemented with 10% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin at 37°C with 5% CO₂. Peripheral blood mononuclear cells (PBMCs) were collected from 4T1 tumor-bearing mice, lysed with an ACK lysis buffer (KD, United States) for 5 min on ice, and then maintained in complete DMEM with 55 µM 2-mercaptoethanol (Gibco, United States).

Tian S., Welte T., Mai J., Liu Y., Ramirez M, & Shen H. (2022). Identification of an Aptamer With Binding Specificity to Tumor-Homing Myeloid-Derived Suppressor Cells. Frontiers in Pharmacology, 12, 752934.

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Cell Culture Conditions for Lung Cancer

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Human lung cancer cells H446 and H446/CDDP cells were cultured in RPMI‐1640 medium (Corning Cellgro, Herndon, VA) supporting with 10% fetal bovine serum (HyClone, Logan, UT), 100 U/mL penicillin, and 100 mg/mL streptomycin (Corning Cellgro) at 37°C with 5% CO₂. 1.0 mg/mL puromycin (Sigma‐Aldrich, St. Louis, Missouri, USA) was additionally used to select and maintain the Trps1 and MGMT stably overexpression cells. HEK‐293T was cultured in DMEM (Corning Cellgro) supporting with 10% FBS (HyClone), 100 U/mL penicillin, and 100 mg/mL streptomycin (Corning Cellgro) at 37°C with 5% CO₂.

Liu H., Liao Y., Tang M., Wu T., Tan D., Zhang S, & Wang H. (2018). Trps1 is associated with the multidrug resistance of lung cancer cell by regulating MGMT gene expression. Cancer Medicine, 7(5), 1921-1932.

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Cell Culture Conditions for Multiple Cell Lines

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MEF, human fibroblast, 293T, M14, SW48, U2OS, and MCF7 cells were cultured in DMEM (Corning, Cellgro) plus 10% fetal bovine serum (FBS) (Gibco), 100 U/mL penicillin G, and 100 μg/mL streptomycin (Corning, Cellgro). A549 and H1299 were cultured in RPMI (Corning, Cellgro) plus 10% FBS, 100 U/mL penicillin G, and 100 μg/mL streptomycin. HCT116 was cultured in McCoy’s 5A (Corning, Cellgro) plus 10% FBS (Gibco), 100 U/mL penicillin G, and 100 μg/mL streptomycin. All cells were cultured in a 37°C, 5% CO₂ incubator.

Liu X.S., Haines J.E., Mehanna E.K., Genet M.D., Ben-Sahra I., Asara J.M., Manning B.D, & Yuan Z.M. (2014). ZBTB7A acts as a tumor suppressor through the transcriptional repression of glycolysis. Genes & Development, 28(17), 1917-1928.

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