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Glutamine penicillin streptomycin

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Glutamine/penicillin/streptomycin is a cell culture media supplement that provides essential nutrients and antimicrobial agents to support the growth and maintenance of cell cultures. It contains the amino acid glutamine, as well as the antibiotics penicillin and streptomycin.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions) Mda mb 231, by American Type Culture Collection (3 mentions) Heparin, by Merck Group (3 mentions) Hydrocortisone, by Merck Group (3 mentions) Singlequot supplements, by Lonza (2 mentions) Fix and perm kit, by Thermo Fisher Scientific (2 mentions) Anti human cd3 pecy7, by Thermo Fisher Scientific (2 mentions) Anti human cd4 alexa fluor 450, by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by Biosera (2 mentions) Brefeldin a, by Merck Group (2 mentions) Ionomycin, by Merck Group (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Mcdb131 media, by Caisson (2 mentions) Ascorbic acid 2 phosphate, by Merck Group (2 mentions) Endothelial cell growth supplement (ecgs), by Biomedical Technologies (2 mentions) Laminin, by Merck Group (2 mentions) Cytosine β d arabinofuranoside ara c, by Merck Group (2 mentions) Heat inactivated fetal bovine serum hi fbs, by Thermo Fisher Scientific (2 mentions)

27 protocols using glutamine penicillin streptomycin

1

Fibroblast Reprogramming Protocol

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Fibroblasts reprogramming was performed following a previously described protocol (Yang et al, 2017, 2019). Briefly, the fibroblasts isolated from dura mater were mixed with episomal vectors expressing Oct4, c‐Myc, Klf4, Sox2 (OCKS 4F, 5 μg) and Rarg, Lfh1 (RL 2F, 5.0 μg), electroporated with Amaxa Nucleofector (Lonza, Germany) and then plated on SNL feeders with M15 media (Knockout DMEM (Invitrogen), 15% foetal bovine serum (Hyclone), 1X glutamine–penicillin–streptomycin (Invitrogen), 1X non‐essential amino acids (Invitrogen)). Upon the appearance of colonies, media was replaced with EPSCM (DMEM/F12 (Invitrogen), 20% knockout serum replacement (Invitrogen), 1X glutamine–penicillin–streptomycin, 1X non‐essential amino acids (Invitrogen), 0.1 mM β‐mercaptoethanol (Sigma), 106 U/ml hLIF (Millipore) supplemented with the following inhibitors: CHI99021 Tocris 1 μM, JNK Inhibitor VIII Tocris 4 μM, SB203580 Tocris 10 μM, A‐419259 Santa Cruz 1 μM and XAV939 Stratech 1 μM). Colonies were isolated and plated in 24‐well SNL feeders’ plates for expansion and characterisation. RNA and DNA were extracted from cell pellets using the RNA/DNA/Protein Purification Plus kit (NORGEN, #47700), following the manufacturer's protocol.
Dumas A.A., Pomella N., Rosser G., Guglielmi L., Vinel C., Millner T.O., Rees J., Aley N., Sheer D., Wei J., Marisetty A., Heimberger A.B., Bowman R.L., Brandner S., Joyce J.A, & Marino S. (2020). Microglia promote glioblastoma via mTOR‐mediated immunosuppression of the tumour microenvironment. The EMBO Journal, 39(15), e103790.
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2

IgG Induction by IL-33 and anti-IgM/CD40L

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Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized peripheral blood using Ficoll-Paque density gradient (GE Healthcare). PBMCs were washed with phosphate-buffered saline (PBS) and cultured in RPMI 1640 medium with 10 % FCS and 1 % glutamine/penicillin/streptomycin (Life Technologies). Recombinant human IL-33 protein was purchased from R&D Systems. Anti-human IgM antibody was purchased from eBioscience. Recombinant human CD40 Ligand (CD40L) was purchased from Life Technologies. For in vitro induction of anti-RBC IgG antibody, PBMCs (1 × 106/ml) were cultured with anti-IgM (10 μg/ml) and CD40L (10 ng/ml) in the presence or absence of IL-33 (0–20 ng/ml). Six days later, the supernatants were collected and assayed for IgG antibodies with Human IgG total ELISA kit (eBioscience).
Bu X., Zhang T., Wang C., Ren T, & Wen Z. (2015). IL-33 reflects dynamics of disease activity in patients with autoimmune hemolytic anemia by regulating autoantibody production. Journal of Translational Medicine, 13, 381.
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3

Culturing Human and Mouse Breast Cancer Cells

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We used MDA-MB-231 and SKBR3 human breast cancer cells (ATCC) and AT-3 breast cancer cells derived from a C57BL/6 mouse (gift of Dr. Abrams, Roswell Park Cancer Institute, Buffalo, NY, USA). Breast cancer cells were cultured in DMEM with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies, Carlsbad, CA, USA). Patient derived glioblastoma multiforme (GBM) primary cells (HF2303 and HF3106) were a gift of Dr. Mikkelsen and Dr. deCarvalho, Henry Ford Hospital, Detroit, MI, USA. Details of the procedure for obtaining these patient-derived neurospheres have been described previously (20 (link)). Briefly, a patient tumor samples tumor samples was cut into 1mm3 pieces, triturated, dissociated, and cultured in neurosphere medium (DMEM/F-12 supplemented with N2 (Life Technologies), 0.5 mg/ml BSA (Sigma Aldrich, St. Louis, MO, USA), 25 µg/ml Gentamicin (Life Technologies), 0.5% antibiotic/antimycotic (Life Technologies), 20 ng/ml bFGF and 20 ng/ml EGF (Peprotech, Rocky Hill, NJ, USA) (21 (link)). GBM cells were maintained in culture for up to passage 20 (low passage) All experiments were performed using cells in exponential growth.
Stacer A.C., Wang H., Fenner J., Dosch J.S., Salomonnson A., Luker K.E., Luker G.D., Rehemtulla A, & Ross B.D. (2015). Imaging Reporters for Proteasome Activity Identify Tumor- and Metastasis-Initiating Cells. Molecular imaging, 14, 414-428.
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4

Culturing Mouse Breast Cancer Cells

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C57BL/6 mouse breast cancer cell lines AT-3 and E0771 (gifts of Dr. Abrams and Dr. Mihich, Roswell Park Cancer Institute) were cultured in DMEM with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies, Carlsbad, CA). We also cultured immortalized mammary fibroblasts from C57BL/6 mice (gift of Dr. Moses, Vanderbilt University) in the same medium. We cultured all cells at 37°C with 5% CO2.
Stacer A.C., Fenner J., Cavnar S.P., Xiao A., Zhao S., Chang S.L., Salomonnson A., Luker K.E, & Luker G.D. (2015). Endothelial CXCR7 Regulates Breast Cancer Metastasis. Oncogene, 35(13), 1716-1724.
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5

Culturing Cells for Cancer Research

Cited 2 times
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We cultured human umbilical vein endothelial cells (HUVEC) in endothelial basal medium with SingleQuot supplements (Lonza) (52 (link)) and other cells in DMEM with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies). We previously described 293T (Open Biosystems) and MDA-MB-231 cells (ATCC) stably transduced with CXCR4 (293T-CXCR4 and 231-CXCR4, respectively) (28 (link), 52 (link)). For animal studies, we used 231-CXCR4 cells that co-express GFP. We used immortalized human mammary fibroblasts (HMF) stably transduced with CXCL12-α fused to Gaussia luciferase (CXCL12-α-GL) or unfused Gaussia luciferase (GL) (26 (link)). We cultured all cells in a 37°C incubator with 5% CO2.
Ray P., Stacer A.C., Fenner J., Cavnar S.P., Meguiar K., Brown M., Luker K.E, & Luker G.D. (2014). CXCL12-γ in Primary Tumors Drives Breast Cancer Metastasis. Oncogene, 34(16), 2043-2051.
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6

Measurement of IL-10 in T Cells

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Cell cultures were performed in RPMI 1640 (Sigma Aldrich, UK) containing 10 % FCS (Biosera, UK) supplemented with glutamine/penicillin/streptomycin (Life Technologies, UK). PBMCs (1 × 106/ml) were stimulated for 4 h with PMA (50 ng/ml; Sigma Aldrich, UK) and Ionomycin (500 ng/ml; Sigma Aldrich, UK) in the presence of Brefeldin A (10 μg/ml; Sigma Aldrich, UK). Post stimulation, cells were washed twice with PBS and stained using anti-human CD3 PEcy7 (eBiosciences, UK; clone: UCHT1) and anti-human CD4 Alexa fluor 450 (eBiosciences, UK; clone: RPA-T4) for 20 min in the dark at 4 °C. Cells were washed and fixed with Reagent A (Fix and Perm kit, Invitrogen, UK) for 30 min in the dark at room temperature. Post incubation, cells were washed and re-suspended in Reagent B (Fix and Perm kit, Invitrogen, UK) and anti-human Alexa fluor 647 IL10 antibody (clone: JES3-9D7) was added to cells that were incubated in the dark at room temperature for 30 min. After washing the cells were resuspended in PBS and analysed on a Cyan ™ ADP (Dako Ltd, UK) and the percentage of CD3+ CD4+ IL10+ T cells and IL10 expression levels (MFI value) by CD4 T cells was recorded.
Duggal N.A., Upton J., Phillips A.C, & Lord J.M. (2015). Development of depressive symptoms post hip fracture is associated with altered immunosuppressive phenotype in regulatory T and B lymphocytes. Biogerontology, 17, 229-239.
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7

Breast Cancer Cell Culture Protocol

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We used AT-3 and E0771 mouse breast cancer cells (gifts of Dr. Abrams and Dr. Mihich, respectively, from Roswell Park Cancer Institute) and immortalized mouse mammary fibroblasts (gift of Dr. Moses, Vanderbilt University). AT-3 adenocarcinoma cells lack estrogen and progesterone receptors; these cells also do not overexpress Her2, which classifies the cells as “triple negative”. E0771 cells are an estrogen-receptor positive medullary adenocarcinoma. We cultured all cells in DMEM (Life Technologies) supplemented with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies). We maintained all cells in a 37°C incubator with 5% CO2.
Fenner J., Stacer A.C., Winterroth F., Johnson T.D., Luker K.E, & Luker G.D. (2014). Macroscopic Stiffness of Breast Tumors Predicts Metastasis. Scientific Reports, 4, 5512.
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8

Culturing Mouse Breast Cancer Cells

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C57BL/6 mouse breast cancer cell lines AT-3 and E0771 (gifts of Dr. Abrams and Dr. Mihich, Roswell Park Cancer Institute) were cultured in DMEM with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies, Carlsbad, CA). We also cultured immortalized mammary fibroblasts from C57BL/6 mice (gift of Dr. Moses, Vanderbilt University) in the same medium. We cultured all cells at 37°C with 5% CO2.
Stacer A.C., Fenner J., Cavnar S.P., Xiao A., Zhao S., Chang S.L., Salomonnson A., Luker K.E, & Luker G.D. (2015). Endothelial CXCR7 Regulates Breast Cancer Metastasis. Oncogene, 35(13), 1716-1724.
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9

Culturing Cells for Cancer Research

Cited 2 times
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We cultured human umbilical vein endothelial cells (HUVEC) in endothelial basal medium with SingleQuot supplements (Lonza) (52 (link)) and other cells in DMEM with 10% fetal bovine serum and 1% glutamine/penicillin/streptomycin (Life Technologies). We previously described 293T (Open Biosystems) and MDA-MB-231 cells (ATCC) stably transduced with CXCR4 (293T-CXCR4 and 231-CXCR4, respectively) (28 (link), 52 (link)). For animal studies, we used 231-CXCR4 cells that co-express GFP. We used immortalized human mammary fibroblasts (HMF) stably transduced with CXCL12-α fused to Gaussia luciferase (CXCL12-α-GL) or unfused Gaussia luciferase (GL) (26 (link)). We cultured all cells in a 37°C incubator with 5% CO2.
Ray P., Stacer A.C., Fenner J., Cavnar S.P., Meguiar K., Brown M., Luker K.E, & Luker G.D. (2014). CXCL12-γ in Primary Tumors Drives Breast Cancer Metastasis. Oncogene, 34(16), 2043-2051.
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10

Isolation and Expansion of Mitral Valve Endothelial Cells

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Clonal VEC populations from mitral valve leaflets from sheep were prepared as described[13 ] and expanded on 1% gelatin-coated dishes in endothelial basal medium-2 (EBM-2 (Lonza, cat # 3156), 10 % heat-inactivated FBS (Hyclone), 1X glutamine/penicillin/streptomycin (Life Technologies, Inc) and 2 ng/ml basic FGF (Roche Applied Science). This medium is referred to as EBM-B. Experiments were performed with mitral VEC clone C4 at passages 10 and 11, mitral VEC clone C5 at passages 9–12 and mitral VEC clone E10 at passages 11 and 12.
Wylie-Sears J., Levine R, & Bischoff J. (2014). Losartan Inhibits Endothelial-to-Mesenchymal Transformation in Mitral Valve Endothelial Cells by Blocking Transforming Growth Factor-β-induced Phosphorylation of ERK. Biochemical and biophysical research communications, 446(4), 870-875.
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