L glutamine

Manufactured by Omega Scientific

Sourced in United States

L-Glutamine is a naturally occurring amino acid that plays a crucial role in various biological processes. It is an important component of protein synthesis and is involved in the metabolism of carbohydrates and fats. L-Glutamine is a key substrate for the production of glutathione, a potent antioxidant, and is also essential for the maintenance of intestinal health.

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Lab products found in correlation

7 protocols using l glutamine

Cryopreserved PBMC Thawing Protocol

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Cryopreserved PBMC were quickly thawed by incubating each cryovial at 37°C for 2 min, and cells transferred into 9 ml of cold medium (RPMI 1640 with L-Glutamine and 25 mM Hepes (Omega Scientific), supplemented with 5% human AB serum (GemCell), 1% Penicillin Streptomycin (Gibco), 1% Glutamax (Gibco)), and 20 U/mL Benzonase Nuclease (Millipore). Cells were centrifuged and resuspended in medium to determine cell concentration and viability using Trypan blue and a hematocytometer. Cells were then kept at 4°C until use for flow cytometry or cell sorting.

Hillman H., Khan N., Singhania A., Dubelko P., Soldevila F., Tippalagama R., DeSilva A.D., Gunasena B., Perera J., Scriba T.J., Ontong C., Fisher M., Luabeya A., Taplitz R., Seumois G., Vijayanand P., Hedrick C.C., Peters B, & Burel J.G. (2023). Single-cell profiling reveals distinct subsets of CD14+ monocytes drive blood immune signatures of active tuberculosis. Frontiers in Immunology, 13, 1087010.

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Glutamine Deprivation Cell Culture Protocol

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For glutamine-deprivation experiments, standard DMEM was removed and cells were washed once with phosphate-buffered saline. Glutamine-free and complete medium were made with DMEM devoid of glutamine, arginine, and lysine (ThermoFisher Scientific, A14431) and retained the same nutrient concentrations as standard DMEM. L-Lysine hydrochloride (Sigma L8662) was added to the medium for a final concentration of 0.8 mM. Complete DMEM was made by supplementing with 2 mM L-Glutamine (Omega Scientific 61015) and 0.5 mM L-Arginine monohydrochloride (Sigma A6969). Glutamine-free DMEM was made with the addition of 0.5 mM L-Arginine monohydrochloride. Arginine-free DMEM was made with the addition of 2 mM L-Glutamine and 0.8 mM L-Lysine. Lysine-free DMEM was made with the addition of 2 mM L-Glutamine and 0.5 mM L-Arginine. All media was supplemented with 10% dFBS (Gemini Bio-Products, Sacramento, CA, USA) and P/S.

Lowman X.H., Hanse E.A., Yang Y., Ishak Gabra M.B., Tran T.Q., Li H, & Kong M. (2019). p53 Promotes Cancer Cell Adaptation to Glutamine Deprivation by Upregulating Slc7a3 to Increase Arginine Uptake. Cell reports, 26(11), 3051-3060.e4.

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Culturing Breast Cancer and Mammary Epithelial Cells

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T47D (ATCC #HTB-133), T47DKBluc (ATCC #CRL-2865) and MCF-7 (ATCC #HTB 22) cells were passaged in growth media containing phenol red–free (PRF) DMEM-F12 (Sigma #D6434) or MEM 1X (Gibco #51200-038) with 10% heat-inactivated FBS (Omega Scientific #FB-02) and

10 μ g / ml

insulin (Sigma #9278),

2 mM

L-glutamine (Hyclone #SH30034.01), gentamycin

15 μ g / ml

(Gibco #15750-060), and 1X antibiotics/antimycotics (AB/AM, Gibco #15240-062) and incubated at 37°C with 5%

{CO}_{2}

. For experiments, cells were grown in clearing media with charcoal-stripped serum (CSS) (MEM 1x with 10% charcoal dextran–treated FBS (Omega Scientific #FB-04),

10 μ g / ml

insulin, and

2 mM

L-glutamine) for

24 - 72 h

before being plated for experiments.
The 76N-Tert cell line, a human mammary epithelial cell line immortalized with expression of human telomerase reverse transcriptase (TERT), was a gift from Dr. Vimla Band (Zhao et al. 2010 (link)). These cells were grown in F-media [

250 mL

DMEM (-pyruvate) (Gibco #11965-092),

250 mL

Ham’s F12 (Gibco #11765-054), 5% FBS,

250 ng / mL

hydrocortisone (Sigma #H4001),

10 ng / mL

human epidermal growth factor (Tonbo Biosciences #21-8356-U100),

8.6 ng / mL

cholera toxin vibrio (Millipore Sigma #227035),

1 μ g / mL

human insulin solution, and 1X antibiotic/antimycotic] and passaged every 2–3 d.

Majhi P.D., Sharma A., Roberts A.L., Daniele E., Majewski A.R., Chuong L.M., Black A.L., Vandenberg L.N., Schneider S.S., Dunphy K.A, & Jerry D.J. (2020). Effects of Benzophenone-3 and Propylparaben on Estrogen Receptor–Dependent R-Loops and DNA Damage in Breast Epithelial Cells and Mice. Environmental Health Perspectives, 128(1), 017002.

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Establishment of EBV-transformed Lymphoblastoid Cell Lines

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EBV–transformed lymphoblastoid B-cell lines (LCL) was developed in our laboratory as described previously [18 ]. Three lines of LCLs were established from patients with EBV+DLBCL (LCL-1), EBV-DLBCL (LCL-2), and a healthy subject (LCL-3), respectively. In brief, peripheral blood samples were collected and used to generate LCL. Concentrated supernatant from B95-8 cultures, a EBV-transformed marmoset B-cell line, was added to parallel samples in the presence of cyclosporin A in RPMI 1640/20% FCS.
SU-DHL-4, SU-DHL-6, HBL-1, and OCI-Ly-10 DLBCL cell lines were gifts from Dr. Ronald Levy (Stanford University, Stanford, CA, USA). Raji and Daudi human Burkitt lymphoma (BL), Karpas 299 anaplastic large cell lymphoma (ALCL) and Jurkat T-cell lymphoblastic leukemia cell lines were obtained from the American Type Culture Collection (ATCC).
SU-DHL-4, HBL-1, LCL, Raji, Daudi, Karpas 299, and Jurkat cells were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) plus 10% heat-inactivated fetal calf serum (FCS; Omega Scientific, Tarzana, CA, USA), 100 U/mL penicillin/streptomycin, and 2 mmol/L L-glutamine, at 37°C in 5% CO₂. SU-DHL-6 cells were cultured in RPMI 1640 medium plus 20% FCS. OCI-Ly-10 cells were cultured in Iscove’s Modified Dulbecco’s Media(IMDM) complete medium plus 20% fresh human plasma (heparinized) instead of FCS.

Quan L., Chen X., Liu A., Zhang Y., Guo X., Yan S, & Liu Y. (2015). PD-1 Blockade Can Restore Functions of T-Cells in Epstein-Barr Virus-Positive Diffuse Large B-Cell Lymphoma In Vitro. PLoS ONE, 10(9), e0136476.

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Pancreatic Cancer Cell Culture Protocol

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The poorly differentiated human pancreatic cancer MIA PaCa-2 cell line and the moderately differentiated BxPC-3 human cell line were obtained from the American Type Culture Collection (Manassas, Va). MIA PaCa-2 cells were cultured in 1/1 D-MEM/F-12 medium (GIBCO Invitrogen Corporation, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS), L-glutamine (final concentration 4mM), and 1% antibiotics/antimycotics solution (Omega Scientific, Tarzana, Calif). BxPC-3 cells were cultured in RPMI-1640 supplemented with 10% FBS and 1% of antibiotic/antimycotics solution. Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂ and were used between passages 2 and 10.

Yang J., Chheda C., Lim A., Hauptschein D., Zayou L., Tang J., Pandol S.J, & Edderkaoui M. (2022). HDAC4 Mediates Smoking-Induced Pancreatic Cancer Metastasis. Pancreas, 51(2), 190-195.

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Pancreatic Cancer Cell Line Cultivation and Transfection

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The poorly differentiated MIA PaCa-2 and moderately differentiated Bx-PC3 human pancreatic adenocarcinoma cell lines were obtained from the American Type Culture Collection (Manassas, VA). MIA PaCa-2 cells were grown in 1/1 D-MEM/F-12 medium (GIBCO Invitrogen Corporation, Grand Island, NY) supplemented with 15% fetal bovine serum (FBS), 4 mM l-glutamine, and 1% of antibiotic/antimicotic solution (Omega Scientific, Tarzana, CA). Bx-PC3 were grown in RPMI-1640 (GIBCO Invitrogen Corporation, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS) and 1% of antibiotic/antimicotic solution (Omega Scientific, Tarzana, CA). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂ and were used between passages 2 and 10.
Transient transfections of MIA PaCa cells were performed using the electroporation Amaxa System Nucleofector^™ (Amaxa Inc, Gaithersburg, MD) according to the manufacturer protocol. HDAC3 siRNA, (MWG biotech, High Point, NC) was applied to the cells using electroporation according to the Amaxa kit protocol. Control cells were transfected with the Silencer Negative Control siRNA #1 (Ambion; Foster City, CA).

Edderkaoui M., Xu S., Chheda C., Morvaridi S., Hu R.W., Grippo P.J., Mascariñas E., Principe D.R., Knudsen B., Xue J., Habtezion A., Uyeminami D., Pinkerton K.E, & Pandol S.J. (2016). HDAC3 mediates smoking-induced pancreatic cancer. Oncotarget, 7(7), 7747-7760.

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Neural Crest Cell Isolation Protocol

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Cultures were prepared roughly following techniques previously described (Bronner-Fraser* and García-Castro, 2008 ; Walheim et al., 2012 ). Chicken eggs were incubated to HH13–17. Embryos were extracted from their yolk into autoclaved Ringer’s where extra-truncal embryonic tissue was removed. Remaining tissue was incubated at 37°C in 5% CO₂ in DMEM-diluted Dispase [0.24 U/ml; Roche (Indianapolis, IN) and Cell Systems (Kirkland, WA)] generally for 75–90 min then placed in L15 for isolation of the NT with fine tools (at times with small amounts of non-NT tissue remaining attached). NTs were cut into segments 1–4 somites long and cultured overnight on a fibronectin-coated surface in DMEM, L-Glutamine (1.8–2 mM), and antibiotics [Penicillin G (90–100 U/ml), Streptomycin (90–100 μg/ml)] (all Omega Scientific; Tarzana, CA), and 8–10% FBS (primarily from Omega Scientific or Life Technologies) to allow for TNCC emigration and halo formation around the NT. In general, cells within resultant TNCC-enriched cultures that exhibited mesenchymal morphology and were not part of the NT were considered TNCCs.

de Bellard M.E., Ortega B., Sao S., Kim L., Herman J, & Zuhdi N. (2018). Neuregulin-1 is a chemoattractant and chemokinetic molecule for trunk neural crest cells. Developmental dynamics : an official publication of the American Association of Anatomists, 247(7), 888-902.

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