The palmitic acid stock solution was prepared using fatty acid-free BSA (A8806, Sigma) at a molar ratio of 3.3:1 (palmitic:BSA) [45 (link)]. To prepare the stock solution, 500 mM palmitic acid (P0500, Merck) was dissolved in absolute ethanol at 70 °C. Then, 10 μL of palmitic acid solution was added to 1 mL prewarmed 10% BSA (prepared in cell culture medium). The mixture was incubated at 55 °C until it became clear and was then filtered before use. For the vehicle control, 10 μL ethanol was mixed with 1 mL 10% BSA. Oleic acid was prepared at a molar ratio of 6:1 (Oleic acid:BSA) using the product (29557, Cayman, MI, USA). The vehicle stock solution was prepared by dissolving 0.8 mM BSA in 150 mM sodium chloride (S3014, Sigma).
Oleic acid
Manufactured by Cayman Chemical
Sourced in United States
Oleic acid is a monounsaturated fatty acid commonly used in various laboratory applications. It serves as a precursor for the synthesis of other organic compounds and is a key component in the formulation of emulsions, lubricants, and personal care products.
Automatically generated - may contain errors
Lab products found in correlation
Linoleic acid, by Cayman Chemical (15 mentions)
Palmitic acid, by Cayman Chemical (8 mentions)
Stearic acid, by Cayman Chemical (5 mentions)
Arachidonic acid, by Cayman Chemical (5 mentions)
Eicosapentaenoic acid, by Cayman Chemical (4 mentions)
Hydrogen peroxide, by Merck Group (3 mentions)
Simvastatin, by Merck Group (3 mentions)
Etomoxir, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Docosahexaenoic acid, by Cayman Chemical (3 mentions)
Palmitic acid, by Merck Group (2 mentions)
Ex 527, by Merck Group (2 mentions)
Compound c, by Merck Group (2 mentions)
N acetylcysteine, by Merck Group (2 mentions)
Mk 2206, by Cayman Chemical (2 mentions)
Allopurinol, by Merck Group (2 mentions)
Diphenyleneiodonium chloride dpi, by Merck Group (2 mentions)
D4902, by Merck Group (2 mentions)
Cerulenin, by Merck Group (2 mentions)
L ng nitroarginine methyl ester l name, by Cayman Chemical (2 mentions)
33 protocols using oleic acid
1
Preparation of Fatty Acid-BSA Solutions
2
Polarization of Bone Marrow-Derived Macrophages
3
Deuterated Fatty Acid Metabolism in Vascular Cells
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Human aorta endothelial cells line (HAEC) was obtained from Lonza (Basel, Switzerland). Vascular smooth muscle cell line (MOVAS) was purchased from American Type Culture Collection (ATCC, Rockville, Maryland, MD, USA). HAECs were maintained in supplemented endothelial growth medium EGM-2 (Lonza, Basel, Switzerland), MOVAS cells were cultured in DMEM supplemented 10% (v/v) FBS (both from Gibco, Scotland, UK). Cultured were maintained at 37 °C in a humidified atmosphere of 5% CO2—95% air. When cells achieved 90% confluence the medium was changed and cells were treated with 100 μM deuterated Oleic acid (OAd34, Sigma Aldrich) or deuterated arachidonic acid (AAd8, 25 µM, Cayman Chemical) in the presence or absence of lipolysis inhibitor, atglistatin (50 µM), phospholipase A2 inhibitor (AACOCF3, 10 µM, Abcam), COX-1 inhibitor (SC-560, Cayman Chemical) or COX-2 inhibitor (Dup-697, Cayman Chemical) for 4 h or 24 h. Oleic acid (Cayman Chemical) for the experiments was freshly saponified using 100 mM NaOH and BSA-conjugated (10% in DPBS, low endotoxin, fatty acid-free, suitable for cell culture, sterile-filtered; Sigma Aldrich). After 4 h or 24 h of incubation cells were fixed for 4 min with 4% paraformaldehyde. Fixed cells were washed 3 times with DPBS and stored in DPBS at 4 °C until the execution of the measurements.
4
Fatty Acids Modulation of Neuronal Activity
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All chemical compounds were purchased from Wako unless otherwise stated. DHA (cis-4,7,10,13,16,19-docosahexaenoic acid: DHA), DHA-Me, ARA, EPA (cis-5,8,11,14,17-eicosapentaenoic acid), DHK, indomethacin, and Triton X-100 were purchased from Sigma. DHA-CoA and CoA were purchased from Avanti. Oleic acid, linOleic acid, ALA, cis-11,14-eicosadienoic acid, cis-11,14,17-eicosatrienoic acid, cis-13,16,19-docosatrienoic acid (DTriA), cis-7,10,13,16-docosatetraenoic acid (DTetA), cis-7,10,13,16,19-docosapentaenoic acid (DPA), and N-(2-hydroxyethyl)-4Z,7Z,10Z,13Z,16Z,19Z-docosahexaenamide (DHA-EA) were purchased from Cayman Chemical. L-Glu stock solution (20 mM) in purified water (DIRECT-Q; Millipore) was diluted to the required final concentration with ND96 just before application. Stock solutions of fatty acids (100 mM) in ethanol was diluted to the required final concentration with ND96 on demand and used within 2 h to avoid precipitation. We confirmed that the vehicle had no effect prior to each experiment.
5
Lipid Metabolism in HepG2 Hepatocellular Carcinoma
6
Fatty Acid Extraction and Analysis
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Phytomonic acid (C18 CFA), palmitoleic acid (C16∶1), palmitic acid (C16∶0), oleic acid (C18∶1n9), and stearic acid (C18∶0) were obtained from Cayman Chemical (MI). Lauric acid (C12∶0), myristic acid (C14∶0), vaccenic acid (C18∶1n7), elaidic acid (trans-C18∶1n9), linoleic acid (C18∶2n6), arachidonic acid (C20∶4n6), and eicosapentaenoic acid (C20∶5n3) were from Nu-Chek Prep, Inc. (MN). All solvents were of GC-MS grade and all other reagents were of the highest grade available.
7
Fatty Acid Modulation of Dendritic Cells
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Fatty acids (FAs) were purchased from Cayman and diluted in ethanol for all FAs [Palmitic acid (PA, C16:0), stearic acid(C18:0), palmitoleic acid (C16:1), oleic acid (OA, C18:1n-9), linoleic acid (LA, C18:2n6c), cis-8,11,14-eicosatrienoic acid (C20:3n6), cis-11,14,17- eicosatrienoic acid (C20:3n3), erucic acid (C22:1n9), eicosapentaenoic acid (EPA, C20:5n3), nervonic acid (C24:1n9), docosahexenoic acid (DHA, C22:6n3) and arachidonic acid (AA, C20:4n-6)] . The FAs-free medium was contained RPMI-1640 basic medium, 1×ITS medium supplement (Sigma) and 20 ng/ml GM-CSF without FAs supplement. Before addition to the culture medium, all FAs were conjugated to bovine serum albumin (BSA, fatty acid-free, Sigma) at the indicated concentration with FAs-free medium containing 1% BSA. The fatty acids were incubated with 1% BSA for 2 h before adding to DC. The concentration of saturated, monounsaturated, and polyunsaturated fatty acids in ITS medium was 20 µM. Half of the medium was discarded and replaced with fresh DC medium every other day. On day 6, DC were infected with the virus for 24 h and then analyzed by FACS.
8
Multiparametric Flow Cytometry Assay
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We followed the procedures described in [33 (link)]. Positive controls cells for LD and apoptosis were treated with 75 µM oleic acid (Cayman Chemical, Ann Arbor, MI, USA) or Staurosporine 10 µM for 24 h, respectively. Cells were then harvested and centrifuged at 300 g, for 10 min at RT. Subsequently, cells were re-suspended in PBS (Sigma-Aldrich) and 5 mM 2′,7′-Dichlorodihydrofluorescein diacetate (CM-H2DCFA) (Thermo Fisher Scientific, St. Louis, MO, USA) (for ROS) or 0.5 µg/mL boron dipyrromethene (BODIPY) 483/503 (for LD) and incubated for 30 min at 37 °C (for ROS) or RT (for LD). Then, 1 drop/0.5 mL of Annexin V-Pacific Blue (Annexin V-Pacific Blue Ready Flow Reagent, Thermo Fisher Scientific, St. Louis, MI, USA) was added to the mixture. After 15 min, the cell fluorescence was measured using the flow cytometer FACS Melody (Becton Dickinson Biosciences, Franklin Lakes, NJ, USA) equipped with violet (405 nm), blue (488 nm) and yellow/green (561 nm) lasers. Cells for ROS positive control were also incubated with 5 mM H2O2 for 30 min after staining and before measurement. For cytotoxicity, we used the Annexin V-Pacific Blue Ready Flow Reagent (Thermo Fisher Scientific, St. Louis, MO, USA) staining kit to monitor apoptosis using flow cytometry. Samples were stained for cytotoxicity when prepared for ROS and LD detection.
9
Fatty Acid Conjugation and Cell Treatment
10
Optimized neuronal culture media
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BrainPhys neuronal media (Stem Cell Technologies) supplemented with 1X B27 with vitamin A (ThermoFisher), 1X N2 Plus media supplement (R&D Systems), 20 ng/ml BDNF (Peprotech), 20 ng/ml GDNF (Peprotech), 1 mM creatine (Sigma-Aldrich), 200 nM L-ascorbic acid (Sigma-Aldrich), 1 μg/ml mouse laminin (ThermoFisher), 0.5 mM glutamax (ThermoFisher), 0.5X penicillin-streptomycin (ThermoFisher), 1X Normocin (Invivogen), 5 ng/ml TGF-b (Peprotech), 100 ng/ml human IL-34 (Peprotech), 1.5 μg/ml cholesterol (Sigma-Aldrich), 1 ng/ml gondoic acid (Cayman Chemicals), 100 ng/ml oleic acid (Cayman Chemicals), 460 μM Thioglycerol (Sigma-Aldrich), 1X Insulin-Transferrin-Selenium (ThermoFisher), 25 ng/ml rhM-CSF (Peprotech), and 5.4 μg/ml Human Insulin Solution (Sigma).
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