Rpmi 1640 l glutamine medium

Manufactured by Thermo Fisher Scientific

Sourced in United States, Belgium

RPMI 1640 + L-glutamine medium is a cell culture medium formulated to support the growth and maintenance of a variety of cell lines. It provides a balanced salt solution, amino acids, vitamins, and other essential nutrients required for cell proliferation. The addition of L-glutamine, a critical nutrient, further enhances the medium's ability to sustain cellular metabolism and growth.

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RPMI 1640 medium (19060 citations) RPMI 1640 (15891 citations) L-glutamine (14671 citations) Glutamine (2902 citations) RPMI medium (1417 citations) 1640 medium (927 citations) RPMI 1640 + L-glutamine (21 citations) 1640 medium (RPMI 1640) (17 citations) RPMI 1640 glutamine [-] medium (3 citations) RPMI Medium 1640 (1×) + L-glutamine (2 citations)

8 protocols using rpmi 1640 l glutamine medium

Stimulation and Cytokine Analysis of Heparinized Blood

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Heparinized blood was diluted with L-glutamine-RPMI 1640 medium (Life
Technologies, Grand Island, NY) at a 1:1 ratio and stimulated with organic dust
extract (1%), triacyl lipopeptide N-palmitoyl-S-dipalmitoylglyceryl
Cys-Ser-(Lys)₄ (Pam3CSK4, 1 ng/ml), peptidoglycan (PGN, 10
µg/ml) or phosphate buffered saline. Blood was incubated for 24 hr at
37°C with 5% CO₂, and then centrifuged at 500
× g for 5 min. Cell-free supernates were stored at −80°C
for later cytokine analysis. Blood samples were processed within 2 hr of
collection.

Smith L.M., Weissenburger-Moser L.A., Heires A.J., Bailey K.L., Romberger D.J, & LeVan T.D. (2017). Epistatic effect of TLR −1, −6 and −10 Polymorphisms on Organic Dust-Mediated Cytokine Response. Genes and immunity, 18(2), 67-74.

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Stimulation and Cytokine Analysis of Heparinized Blood

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Cell Culture Protocols for Skin and Lymphoma

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Sézary syndrome cell line HuT78 was a gift from Michael U. Martin (Justus-Liebig-Universität Gießen, Gießen, Germany), mycosis fungoides cell line MyLa was a gift from Jean-Philippe Merilo and Edith Chevret (University of Bordeaux, Bordeaux, France), HaCaT cells were a gift from Norbert Fusenig (German Cancer Research Institute, Heidelberg, Germany). HuT78, and MyLa cells were grown in RPMI 1640 medium [-] L-Glutamine with 1% penicillin/streptomycin, 1% L-Glutamine (all Gibco/Thermo Fisher, Waltham, MA, USA), 10% fetal calf serum (FCS) (Sigma-Aldrich, St. Louis, MO, USA). HaCaT cells in DMEM, high glucose, GlutaMAX^TM (Gibco/Thermo Fisher, Waltham, MA, USA) with 1% penicillin/streptomycin, 10% FCS. Normal primary human keratinocytes were isolated from infantile foreskins of donors and seeded in DermaLife^® K cell medium (CellSystems, Troisdorf, Germany). All cells were incubated at 37 °C in a humidified 5% CO₂ incubator.

Wilhelm R., Eckes T., Imre G., Kippenberger S., Meissner M., Thomas D., Trautmann S., Merlio J.P., Chevret E., Kaufmann R., Pfeilschifter J., Koch A, & Jäger M. (2021). C6 Ceramide (d18:1/6:0) as a Novel Treatment of Cutaneous T Cell Lymphoma. Cancers, 13(2), 270.

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INS-1E Cell Culture Protocol

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INS-1E cells were cultured in a RPMI-1640 medium (+l-Glutamine, Gibco, Ghent, Belgium) supplemented with 10% fetal bovine serum (FBS, Hyclone, Aalst, Belgium), 10 mM Hepes (Sigma, Bornem, Belgium), 1 mM sodium-pyruvate (Gibco), 0.05 mM 2-mercaptoethanol (Gibco), and 1% penicillin/streptomycin (Cellgro, Manassas, WA, USA).

Ribeiro R.S., Ketkar-Atre A., Yin T., Louchami K., Struys T., Lambrichts I., Sener A., Malaisse W.J., De Cuyper M, & Himmelreich U. (2018). Improved Labeling of Pancreatic Islets Using Cationic Magnetoliposomes. Journal of Personalized Medicine, 8(1), 12.

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Colon-26 Adenocarcinoma Cell Culture

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Colon-26 adenocarcinoma (C26) cells (Division of Cancer Treatment and Diagnosis Tumor Repository, National Cancer Institute) were cultured in RPMI 1640 + L-glutamine medium (Gibco, Thermo Fisher Scientific) supplemented with 5% (v/v) fetal bovine serum (FBS) and 1% (v/v) Penicillin–Streptomycin (10,000 U/ml) at 37 °C with 5% CO₂. At ~ 75% confluence, cells were washed, trypsinized, centrifuged, and resuspended in sterile PBS at a concentration of 1.0 × 10⁷ cells/ml immediately prior to mouse inoculation.

Law M.L, & Metzger J.M. (2021). Cardiac myocyte intrinsic contractility and calcium handling deficits underlie heart organ dysfunction in murine cancer cachexia. Scientific Reports, 11, 23627.

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Liver Cell Culture and Transfection

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HepG2.2.15 cells, HepG2 cells, and Huh7 cells were cultured at 37 °C in a CO₂ incubator. HepG2.2.15 cells were routinely cultured in RPMI-1640 medium (Gibco, America) supplemented with 10% inactivated fetal bovine serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco, America), 1% nonessential amino acids (NEAA), 1% HEPES, and 500 μg/mL G418 (Gibco, America). HepG2 cells and Huh7 cells were cultured in DMEM medium, supplemented with 10% inactivated FBS, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco, America), 1% NEAA, and 1% HEPES. The transfection of siRNAs and plasmids was performed with Lipofectamine 2000 transfection reagent (Invitrogen, America) according to the manufacturer’s instructions. Peripheral blood mononuclear cells (PBMCs) from CHB patients (obtained with signed informed consent) were separated by Ficoll density gradient centrifugation and cultured in RPMI-1640 + L-glutamine medium (Gibco, America) supplemented with 10% inactivated FBS. DOTAP liposomal transfection reagent (Sigma-Aldrich) was used in the transfection of PBMCs with siRNAs.

Lan T., Wei Z., He Y., Wan S., Liu L., Cheng B., Li R., Chen H., Liu G, & Meng Z. (2021). Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity. Virology Journal, 18, 37.

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Syngeneic 4T1 Breast Cancer Model

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4T1 cell line (ATCC) was tested for mycoplasma (by PCR using standard mycoplasma testing protocol) and cultured in RPMI-1640 [+] L-glutamine medium (Gibco, #11875-093) supplemented with 100 IU/ml penicillin and 100 μg/ml streptomycin (Lonza, # DE17-602E) and 10% heat-inactivated fetal bovine serum (FBS, Sigma, #F2442). Female Balb/c mice were purchased from Charles River Laboratories (Wilmington, MA, USA) and kept under pathogen-free conditions at the National Cancer Institute Animal Facility in Frederick.
The studies were approved by the National Cancer Institute-Frederick Animal Care and Use Committee. NCI-Frederick is accredited by AAALAC International and follows the Public Health Service Policy for the Care and Use of Laboratory Animals. Animal care was provided in accordance with the procedures outlined in the “Guide for Care and Use of Laboratory Animals (National Research Council; 1996; National Academy Press; Washington, D.C.).

Stravokefalou V., Stellas D., Karaliota S., Nagy B.A., Valentin A., Bergamaschi C., Dimas K, & Pavlakis G.N. (2023). Heterodimeric IL-15 (hetIL-15) reduces circulating tumor cells and metastasis formation improving chemotherapy and surgery in 4T1 mouse model of TNBC. Frontiers in Immunology, 13, 1014802.

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LNCaP Cell Fixation and Chromatin Preparation

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The androgen-sensitive human prostate adenocarcinoma cell line LNCaP was purchased from the American Type Culture Collection (ATCC) and cultured in RPMI 1640 (+) L-Glutamine medium from Gibco Life Technologies (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% fetal bovine serum, 100 units/ml penicillin-streptomycin, and mM sodium pyruvate. This line was authenticated regularly in our institutional CCSG Cell Line Characterization Core and also examined to be free of mycoplasma contamination. Cells were maintained in 5% CO2 at 37C and cultured in 10 cm plates to a confluency of 70-80% before fixation. LNCaP cells were fixed in 1% methanol-free formaldehyde (Thermo Fisher Scientific) in RPMI for 10 min at room temperature followed by quenching for 5 min in mM glycine (Sigma, St. Louis, MO) with low-speed shaking on an orbital platform. Plates were washed 2× with ice-cold phosphate-buffered saline (PBS) (pH 7.4) (Thermo Fisher Scientific) to eliminate any residual media, and 7 ml cold PBS containing 1× Complete Protease Inhibitors Cocktail, EDTA free (Roche, Basel, Switzerland) were immediately added. Plates were kept on ice while the cells were harvested by scraping. Cells were transferred to 15 ml conical tubes (2 plates/tube) and collected by centrifugation (4 min at 805 × g at 4°C). Cells were immediately used for chromatin preparation.

Simper M., L D.C., Gaddis S., Lin K., Mikulec C., Takata Y., Tomida M., Zhang D., Tang D., Estecio M., Shen J., & Lu Y. (2022). Commercial ChIP-Seq library preparation kits performed differently for different classes of protein targets.

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