African green monkey kidney epithelial cells (VeroE6) were from Biomedica (VC-FTV6, Vienna, Austria) and grown in Minimal Essential Medium (MEM) containing Earle’s Salts and L -Glutamine (Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 2% fetal calf serum (FCS) (Thermo Fisher Scientific) and 1% Penicillin-Streptomycin (PenStrep) (Thermo Fisher Scientific).
Earle s salts and l glutamine
Manufactured by Thermo Fisher Scientific
Sourced in United States
Earle's salts and L-glutamine are commonly used cell culture media supplements. Earle's salts provide a balanced salt solution to support cell growth and maintain osmotic pressure, while L-glutamine is an essential amino acid required for cell metabolism. These products are intended for research use only and their core function is to support in vitro cell culture applications.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Hepes buffer, by Thermo Fisher Scientific (2 mentions)
Pen strep, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (2 mentions)
Insulin transferrin selenium, by Thermo Fisher Scientific (2 mentions)
Earle s salt, by Thermo Fisher Scientific (2 mentions)
2 3 butanedione monoxime, by Thermo Fisher Scientific (2 mentions)
Laminin, by Corning (2 mentions)
Nonessential amino acid, by Thermo Fisher Scientific (1 mentions)
Sodium tripolyphosphate, by Merck Group (1 mentions)
Acetic acid, by Avantor (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2 h tetrazolium bromide mtt, by Thermo Fisher Scientific (1 mentions)
Methanol, by Avantor (1 mentions)
14 protocols using earle s salts and l glutamine
1
Culturing VeroE6 Cells for Research
2
Culturing THP-1, HeLa, and Ehrlichia chaffeensis
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Human monocytic leukemia cells (THP-1; ATCC TIB-202) were propagated in RPMI 1640 with L-glutamine and 25 mM HEPES buffer (Invitrogen, Carlsbad, CA), supplemented with 10% fetal bovine serum, and incubated at 37°C in a 5% CO2 atmosphere. Henrietta Lack’s cervical epithelial adenocarcinoma (HeLa) cells were propagated in MEM medium with Earle’s Salts and L-glutamine (Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum. E. chaffeensis (Arkansas strain) was cultivated in THP-1 cells as previously described (Byerly et al., 2022 (link)).
3
Curcumin-Chitosan Nanoparticles for FIPV
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Curcumin powder, low-molecular weight chitosan (75–85% deacetylated), Tween 80, sodium tripolyphosphate (TPP), sucrose, and sodium hydroxide (NaOH) were purchased from Sigma-Aldrich (St Louis, MO, USA). Acetic acid, ethyl acetate, and methanol were purchased from AMRESCO (Radnor, PA, USA). Absolute ethanol was purchased from Scharlau (Barcelona, Spain). All reagents utilised in this study were of analytical grade, except methanol, which was high-performance liquid chromatography (HPLC) grade. CrFK cells and FIPV strain WSU 79-1146 were purchased from ATCC (Manassas, VA, USA). Minimum essential medium (MEM) containing Earle's salts and L-glutamine, foetal bovine serum, nonessential amino acid (NEAA), penicillin (100 U/mL), streptomycin (100 μg/mL), phosphate-buffered saline (PBS), TryPLE™, trypan blue, dimethyl sulfoxide (DMSO), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
4
Cultivation of THP-1 and HeLa Cell Lines
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Human monocytic leukemia cells (THP-1) were grown and maintained in RPMI medium 1640 with L-glutamine and 25 mM HEPES buffer (Invitrogen, Carlsbad, CA), supplemented with 10% fetal bovine serum (HyClone, Logan, UT). Henrietta Lack’s cervical epithelial adenocarcinoma (HeLa) cells were propagated in MEM medium with Earle’s Salts and L-glutamine (Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum (HyClone). E. chaffeensis (Arkansas strain) was cultivated in THP-1 cells as previously described [77 (link)].
5
Culturing and Differentiating Leishmania Parasites
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L. mexicana (World Health Organization strain MNYC/BZ/62/M397) promastigotes were grown in M199 medium with Earle's salts and L-glutamine (Thermo Fisher), supplemented with 10% foetal calf serum (FCS) (Thermo Fisher), 5 mM HEPES·NaOH (pH 7.4), 26 mM NaHCO3 and 5 µg/ml haemin, at 28°C. Cells were maintained in logarithmic growth at culture densities between 1×105 cells/ml and 1×107 cells/ml through regular subculture (Wheeler et al., 2011 (link)). Culture densities were measured with a CASY model TT cell counter (Roche Diagnostics). Axenic amastigotes were generated by subculturing into Schneider's Drosophila medium (Thermo Fisher) supplemented with 20% FCS (Thermo Fisher) and 25 mM MES·HCl (pH 5.5) at 34°C under 5% CO2 (Bates, 1994 (link)), and growth for 72 h without subculture. Amastigotes were generated by infection of J774 macrophages with stationary-phase promastigotes and allowed to differentiate into amastigotes for 72 h. J774 macrophages were grown in RPMI (Thermo Fisher), supplemented with 10% FCS (Thermo Fisher), at 37°C (prior to infection) or at 34°C (after infection) under 5% CO2.
6
Culturing Human Bronchial Epithelial Cells
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Human bronchial epithelial cells obtained from lung tissues removed during surgery and cultured at an air-liquid interface (ALI) were purchased from Epithelix (Plan-les-Ouates, Switzerland) [23] . Cells were cultured with MucilAir™ culture medium (Epithelix) for at least 8 wk. The permeability of tight junctions and the differentiation were validated by transepithelial electrical resistance measurement (>400 .cm²) and cilia beating frequency (>7 Hz). Data were generated from human bronchial epithelial cells from five CF patients homozygous for the F508del mutation and five healthy donors (non-CF). The human bronchial epithelial cell lines CFBE41o - (CF) and 16HBE14o -(non-CF) were a gift from Prof. D.C. Gruenert (UCSF, San Francisco, USA). They were cultured in minimum essential medium (MEM) in the presence of Earle's salts and L-glutamine (Thermo Fisher, Villebon-sur-Yvette, France) containing 10% bovine growth serum (Eurobio, les Ulis, France) and 1% penicillin/streptomycin (Thermo Fisher). Cell cultures were grown and maintained at 37 °C in a 5% CO 2 humidified incubator.
7
Crocin Quantification in Saffron Extract
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Zolpidem tartrate BP was purchased from Ipca Co. 10×Hank’s balanced salt solution (HBSS), 1 M HEPES buffer, minimal essential medium (MEM) with Earle’s salts and L-glutamine, penicillin-streptomycin, fetal bovine serum (FBS), neuro basal medium, glutaMAX-I supplement, B27 serum-free supplement, trypsin, cell-freezing medium from was purchased from Invitrogen. Acrylamide, ᴅ-glucose, sodium pyruvate, putrescine, progesterone, selenium dioxide, bovine transferrin, insulin from Sigma. Crocin was quantified through the authentic method in an aqueous saffron extract [36 (link)]. Crocin was 95% pure.
8
Bovine Oocyte Maturation Protocol
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Bovine ovaries of zebu breeds were collected from local slaughterhouses, transported to the laboratory in a saline solution (0.9%) at 35°C and processed within 4 hr. Cumulus-oocyte complexes (COCs; n = 2,767, five replicates per treatment) were aspirated from follicles between 3 and 8 mm in diameter and stored in 15 ml conical polystyrene tubes, in a 36°C water bath for 10 min before decantation.
For each treatment, groups of 30-35 COCs with a homogeneous cytoplasm and several cell layers were selected (Stojkovic et al., 2001) and cultured for in vitro maturation in TCM-199 with Earle's salts and l-glutamine (Gibco ® ; Invitrogen Co., Grand Island, NY, USA) supplemented with 10% foetal bovine serum (v/v), 0.2 mM pyruvate, 5 mg/ml luteinizing hormone (Lutropin-V ® ; Bioniche Co., Belleville, ON, Canada), 1 mg/ml follicle-stimulating hormone (Folltropin ® ; Bioniche Co.), 75 μg/ml amikacin and 1 mM cystamine. The COCs were matured in drops (200 μl) covered with mineral oil (Corning, NY, USA) for 22-24 hr at 38.5°C in 5% CO 2 in air and saturated relative humidity without condensation.
For each treatment, groups of 30-35 COCs with a homogeneous cytoplasm and several cell layers were selected (Stojkovic et al., 2001) and cultured for in vitro maturation in TCM-199 with Earle's salts and l-glutamine (Gibco ® ; Invitrogen Co., Grand Island, NY, USA) supplemented with 10% foetal bovine serum (v/v), 0.2 mM pyruvate, 5 mg/ml luteinizing hormone (Lutropin-V ® ; Bioniche Co., Belleville, ON, Canada), 1 mg/ml follicle-stimulating hormone (Folltropin ® ; Bioniche Co.), 75 μg/ml amikacin and 1 mM cystamine. The COCs were matured in drops (200 μl) covered with mineral oil (Corning, NY, USA) for 22-24 hr at 38.5°C in 5% CO 2 in air and saturated relative humidity without condensation.
9
Isolation and Culturing of Murine Ventricular Cardiomyocytes
10
Isolation and Culturing of Murine Ventricular Cardiomyocytes
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Mice ventricular myocytes were isolated by enzymatic digestion using a Langendorff perfusion apparatus77 (link). Cardiomyocytes were isolated from 8–12 week-old non-transgenic (C57BL/6) and transgenic mice. After isolation, the cells were resuspended in perfusion solution with fetal bovine serum (FBS, 5%) and calcium was added gradually to a final concentration of 0.5 mM. Cells were plated on laminin (Corning) coated glass coverslips initially in ‘plating’ media composed of MEM medium with Earle’s salts and L-glutamine (Gibco) plus 5% FBS, 1% penicillin/streptomycin, and 10 mM 2,3-butanedione monoxime (BDM). Cells were allowed to adhere for 2 hour before switching to ‘maintenance’ media containing MEM with Earle’s salts, L-glutamine, 1% penicillin–streptomycin, bovine serum albumin (0.5 mg/ml), 10 mM BDM, 1% insulin-transferrin-selenium (Gibco), 5 mM creatine, 5 mM taurine, 2 mM L-carnitine, 25 μM Blebbistatin. For manipulation of FHF levels, 1 – 2 μL of relevant adenovirus was resuspended in culture medium and added to cells. The ‘maintenance’ media was replaced 24 h to reduce potential toxicity from adenovirus. The efficacy of viral transduction was assessed 24–48 h post infection.
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