Glutamine solution

Manufactured by Thermo Fisher Scientific

Sourced in United States

Glutamine solution is a liquid laboratory product that provides a source of the amino acid glutamine. Glutamine is a key nutrient required for cell growth and proliferation in cell culture applications.

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Glutamine (2902 citations) L-glutamine solution (33 citations) Penicillin-streptomycin-glutamine solution (27 citations) 100× penicillin–streptomycin–l-glutamine solution (5 citations) Penicillin‐streptomycin‐glutamine (100×) solution (3 citations) Penicillin-streptavidin-glutamine-solution (2 citations) Pen/strep/glutamine solution (2 citations) Gentamicin–glutamine solution (2 citations)

7 protocols using glutamine solution

Cell Culture Conditions for Diverse Cell Lines

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Murine T-lymphoma ^MYC-Tet-Off cells (female) were grown in RPMI medium (Thermo Fisher Scientific) supplemented with 10% FBS (Sigma), 1% penicillin/streptomycin solution (Sigma), 1% glutamine solution (Thermo Fisher Scientific), 1% MEM non-essential amino acids (Thermo Fisher Scientific), and 50 μM β-mercaptoethanol (Sigma). Human U2OS (female), U2OS ^MYC-Tet-On, HEK293 (female) and HeLa (female) cells, as well as murine NIH 3T3 (not applicable) cells, were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% FBS (Sigma) and 1% penicillin/streptomycin solution (Sigma). Human mammary epithelial cells (HMLE, female) were cultivated in advanced DMEM/F12 (Thermo Fisher Scientific) supplemented with 20 ng/ml EGF (human) (Thermo Fisher Scientific), 0.5 μg/ml Hydrocortisone (Sigma), 10 μg/ml Insulin (human) (Sigma), 1% penicillin/streptomycin solution (Sigma), 1% glutamine solution (Thermo Fisher Scientific) and 15 mM HEPES (pH 7.2). Human Neuroblastoma SH-EP cells (female) were verified by STR profiling and grown in RPMI medium (Thermo Fisher Scientific) supplemented with 10% FBS and 1% penicillin/streptomycin solution. All cell lines were cultured at 37°C, 5% CO₂ and routinely screened and found negative for mycoplasma contamination in a PCR-based assay.

Baluapuri A., Hofstetter J., Dudvarski Stankovic N., Endres T., Bhandare P., Vos S.M., Adhikari B., Schwarz J.D., Narain A., Vogt M., Wang S.Y., Düster R., Jung L.A., Vanselow J.T., Wiegering A., Geyer M., Maric H.M., Gallant P., Walz S., Schlosser A., Cramer P., Eilers M, & Wolf E. (2019). MYC Recruits SPT5 to RNA Polymerase II to Promote Processive Transcription Elongation. Molecular Cell, 74(4), 674-687.e11.

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Generation and Differentiation of ER-Hoxb8 Cells

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ER-Hoxb8 cells were generated as described earlier (Wang et al., 2006a (link)) and grown in RPMI medium (Thermo Fisher Scientific) supplemented with 10% FBS (Biowest), 1% penicillin/streptomycin solution (Sigma-Aldrich), 1% glutamine solution (Thermo Fisher Scientific), 40 ng/ml recombinant mouse GM-CSF (ImmunoTools) an 1 µM β-estradiol (Sigma-Aldrich). For differentiation, precursor cells were washed and incubated in estradiol-free medium containing 40 ng/ml GM-CSF for several days. HEK293T were grown in DMEM (Thermo Fisher Scientific) supplemented with 10% FBS (Biowest) and 1% penicillin/streptomycin solution (Sigma-Aldrich), 1% glutamine solution (Thermo Fisher Scientific), and 1% sodium pyruvate (Merck). All cell lines were cultured at 37°C, 5% CO₂, and routinely screened and found negative for mycoplasma contamination in a PCR-based assay (PromoCell).

Jauch-Speer S.L., Herrera-Rivero M., Ludwig N., Véras De Carvalho B.C., Martens L., Wolf J., Imam Chasan A., Witten A., Markus B., Schieffer B., Vogl T., Rossaint J., Stoll M., Roth J, & Fehler O. (2022). C/EBPδ-induced epigenetic changes control the dynamic gene transcription of S100a8 and S100a9. eLife, 11, e75594.

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Culturing Chicken Macrophage HTC Cells

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A naturally transformed line of chicken macrophages named HTC cells [22 (link)] were cultured in Roswell Park Memorial Institute (RPMI) 1640 media (Thermo Fisher Scientific, Carlsbad, CA) containing 10% fetal bovine serum (Thermo Fisher Scientific), 1X antibiotic antimycotic solution (Sigma-Aldrich, St Louis, MO, USA), 1X sodium pyruvate solution (Sigma-Aldrich), gentamicin solution (Sigma-Aldrich), 10 mM glutamine solution (Thermo Fisher Scientific) at 37°C for 24–48 h in a humidified incubator containing 5% CO₂ as described earlier with minor modifications. The cells were cultured to semi-confluence followed by dissociation with Accumax (Sigma-Aldrich) to perform different assays.

Gupta A., Bansal M., Liyanage R., Upadhyay A., Rath N., Donoghue A, & Sun X. (2021). Sodium butyrate modulates chicken macrophage proteins essential for Salmonella Enteritidis invasion. PLoS ONE, 16(4), e0250296.

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Generation of Polarized Murine Macrophages

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BMDMs were obtained by flushing the femurs from WT and C/EBPδ KO mice. Erythrocytes were depleted by osmotic shock. Cells were washed and separated using a Ficoll gradient (PAN-Biotech). Primary monocytes (M₀) were generated by culturing for 3 days in DMEM containing with 10% FBS (Biowest) and 1% penicillin/streptomycin solution (Sigma-Aldrich), 1% glutamine solution (Thermo Fisher Scientific), and 15% of L929 supernatant as a source of macrophage colony-stimulating factor. Stimulation of cells with 50 ng/ml IFN-γ (ImmunoTools) and 10 ng/ml LPS (Sigma-Aldrich) for 24 hr was used for M₁-polarization, whereas a 24 hr stimulation with 20 ng/ml IL-4 (Peprotech) led to M₂-polarization of BMDMs.

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Isolation and Culture of Mouse Hippocampal Neurons

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Mouse hippocampal neurons were isolated from postnatal day 1 (P1) C57BL/6J mice, as described [23 (link)], with slight modifications. Briefly, the hippocampus was dissected in Hank’s balanced salt solution (HBSS) and incubated at 37 °C for 15 min with trypsin/ethylenediaminetetraacetic acid (EDTA) (Sigma-Aldrich, St. Louis, MO, USA). After 3 washes in HBSS, tissue was triturated using a sterile 9-inch Pasteur pipette. HBSS was replaced with Neurobasal plating medium (neurobasal medium, Gibco) containing B27 supplement (1:50) (Gibco), 0.5-mM glutamine solution (Gibco), penicillin/streptomycin (Gibco), 1-mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (Hyclone), and 10% heat-inactivated donor horse serum (Gibco). Neuroblasts were plated on poly-D-lysine-coated glass coverslips (p96) at a density of 3 × 10⁴ cells/well and placed in a 37 °C, 5% CO₂ incubator overnight. Next day, in vitro neurobasal plating medium was replaced with neurobasal feeding medium (neurobasal medium containing B27 supplement (1:50), 0.5-mM glutamine solution, penicillin/streptomycin (1:200), and 1-mM HEPES). Half of the feeding medium was replaced with fresh medium every 4 days.

Bobadilla M., García-Sanmartín J, & Martínez A. (2021). Natural Food Supplements Reduce Oxidative Stress in Primary Neurons and in the Mouse Brain, Suggesting Applications in the Prevention of Neurodegenerative Diseases. Antioxidants, 10(1), 46.

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Isolation and Sorting of Monocyte Subsets

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Peripheral blood mononuclear cells were isolated by gradient centrifugation (1.077 g/ml Pancoll, PAN Biotech) from fresh EDTA blood or buffy coats (German Red Cross Blood Transfusion Service, Berlin) of healthy donors, followed by immunomagnetic depletion of CD3⁺/CD19⁺/CD20⁺/CD56⁺/CD235a⁺ cells using biotinylated antibodies (Biolegend) and MagniSort Streptavidin Negative Selection Beads (Invitrogen) (Key resources table). Subsequently monocyte subsets were sorted using a BD FACSAria SORP cell sorter (BD Biosciences) starting with HLA-DR⁺, CD3^-/CD19^-/CD20^-/CD56^- cells following diverse gating strategies: classical monocytes (CD14⁺, CD16^-), non-classical monocytes (CD14^dim CD16⁺), myeloid dendritic cells (cDC2) (CD14^-/CD16^-/CD141^-/CD304^-, CD1c⁺), and plasmacytoid dendritic cells (CD14^-/CD16^-/CD141^-/CD1c^-, CD304⁺). Cells were washed in RPMI 1640 (GIBCO) supplemented with 10% (v/v) FCS (Sigma), 1% (v/v) non-essential amino acid solution (Sigma), 1% (v/v) HEPES (Sigma), 1% (v/v) Glutamine solution (GIBCO) and 1% (v/v) sodium pyruvate (GIBCO).

Wendisch D., Dietrich O., Mari T., von Stillfried S., Ibarra I.L., Mittermaier M., Mache C., Chua R.L., Knoll R., Timm S., Brumhard S., Krammer T., Zauber H., Hiller A.L., Pascual-Reguant A., Mothes R., Bülow R.D., Schulze J., Leipold A.M., Djudjaj S., Erhard F., Geffers R., Pott F., Kazmierski J., Radke J., Pergantis P., Baßler K., Conrad C., Aschenbrenner A.C., Sawitzki B., Landthaler M., Wyler E., Horst D., Hippenstiel S., Hocke A., Heppner F.L., Uhrig A., Garcia C., Machleidt F., Herold S., Elezkurtaj S., Thibeault C., Witzenrath M., Cochain C., Suttorp N., Drosten C., Goffinet C., Kurth F., Schultze J.L., Radbruch H., Ochs M., Eils R., Müller-Redetzky H., Hauser A.E., Luecken M.D., Theis F.J., Conrad C., Wolff T., Boor P., Selbach M., Saliba A.E, & Sander L.E. (2021). SARS-CoV-2 infection triggers profibrotic macrophage responses and lung fibrosis. Cell, 184(26), 6243-6261.e27.

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CaCo-2 Cell Culture Protocol

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CaCo-2 cells were obtained from Health Protection Agency Culture Collections (Salisbury, UK: order no. 86010202). The genotype (STR profiles according to ATCC documentation; CaCo-2 HTB-37) had been analyzed at the start of this study.
The cells were cultured in modified minimum essential Eagle medium (MEM) according to Sigma (product no. M2279, Buchs, CH), with different concentrations of heat-inactivated FCS (1, 5 and 10%) (Lonza, Verviers, B; Cat. No. DE14-801FH, Lot No. 9SBO22H2), 1% penicillin–streptomycin–neomycin-solution (PSN, Gibco, Life Technologies, Basel, CH), 1% glutamine solution (Gibco, Life Technologies), 1% non-essential amino acid solution (Sigma), 1 mM sodium pyruvate (Gibco, Life Technologies) and 1% vitamin solution (Sigma) under uniform cell culture conditions (5% CO₂, 95% humidity and 37 °C). The cultures were passaged once weekly before use in the experiments.

Kaiser J.P., Roesslein M., Diener L., Wichser A., Nowack B, & Wick P. (2017). Cytotoxic effects of nanosilver are highly dependent on the chloride concentration and the presence of organic compounds in the cell culture media. Journal of Nanobiotechnology, 15, 5.

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