The ultra-high-performance liquid chromatography and the LTQ Orbitrap XL mass spectrometry instruments were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Acetonitrile (chromatographic purity) and formic acid (chromatographic purity) were purchased from Merck (Germany). The standards including lysoPC (16:0), lysoPC (17:0), lysoPC (18:0) and taurocholate were purchased from Sigma-Aldrich (USA). Distilled water was filtered by the Milli-Q system (Millipore, USA) before use.
Lysopc
Manufactured by Merck Group
Sourced in United States
LysoPC is a laboratory product manufactured by Merck Group. It is a phospholipid compound used in various scientific research and analytical applications. The core function of LysoPC is to serve as a research tool for the study of lipid metabolism and signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Merck Group (4 mentions)
Human plasminogen, by Enzyme Research (3 mentions)
Acetonitrile, by Merck Group (2 mentions)
Monoclonal antibody for α smooth muscle actin, by Agilent Technologies (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Sprague dawley rat, by Charles River Laboratories (2 mentions)
Caffeine, by Thermo Fisher Scientific (1 mentions)
Ultramark 1621, by Thermo Fisher Scientific (1 mentions)
Accela system, by Thermo Fisher Scientific (1 mentions)
Ltq orbitrap xl hybrid mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Milli q reagent water system, by Merck Group (1 mentions)
Adenosine monophosphate, by Merck Group (1 mentions)
Lysope, by Merck Group (1 mentions)
Nicotinamide, by Merck Group (1 mentions)
Galactosylceramide, by Merck Group (1 mentions)
Cellstar tissue culture flasks, by Greiner (1 mentions)
Sphingosine 1 phosphate (s1p), by Merck Group (1 mentions)
Millipore water purification system, by Merck Group (1 mentions)
L citrulline, by Merck Group (1 mentions)
11 protocols using lysopc
1
Lipid Profiling by UHPLC-Orbitrap
2
UPLC-MS Analysis of Lysophosphatidylcholines
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All solvents used in the present study were high performance liquid chromatography (HPLC) grade without modification. Formic acid and acetonitrile were obtained from Merck (Merck Millipore, Darmstadt, Germany). Distilled water was produced using a Milli-Q Reagent Water System (EMD Millipore, Billerica, MA, United States). Standard preparations of Lysophosphatidylcholine (LysoPC) (16:0) and LysoPC (18:0) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Calibration standards [caffeine, Ultramark 1621 and methionine-arginine-phenylalanine-alanine (MRFA)] were provided by Thermo Fisher Scientific Inc. (Waltham MA, United States). UPLC was performed using an Accela system (Thermo Fisher Scientific Inc. Waltham, MA, United States). MS was performed with a LTQ Orbitrap XL hybrid mass spectrometer (Thermo Fisher Scientific Inc.).
3
Quantification of Lipid Metabolites
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High-performance liquid chromatography grade solvents, including methanol, acetonitrile, and water, were purchased from J.T. Baker (PA, USA). Formic acid, nicotinamide, adenosine monophosphate, LysoPC, and LysoPE were obtained from Sigma-Aldrich (MO, USA) and Avanti Polar Lipids (AL, USA).
4
Lipid Standards for Metabolomics Analysis
5
Preparation and Characterization of Scue Compound
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Scu was purchased from Mianning Jiexiang Co. Ltd. (Chengdu, China). Scue was prepared according to our previously reported methods [28 (link), 29 (link)]. Nimodipine was purchased from Central Pharmaceutical Co. Ltd. (Tianjin, China). Acetonitrile and methanol were of HPLC grade and purchased from Tedia (Fairfield, OH, USA). Analytical grade formic acid was purchased from Merck (Darmstadt, Germany). Deionized water was purified using a Millipore water purification system (Millipore, Milford, MA, USA) and filtered with 0.22-μm membranes. All other reagents used were of analytical grade. l -Aspartic acid, l -citrulline, and lysoPC (18:0) were obtained from Sigma-Aldrich (St. Louis, MO, USA).
6
Calcium Signaling and Apoptosis Assays
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LysoPC (#L1381), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, #M2128), nifedipine (#7634), KB-R7943 (#K4144), LaCl3 (#449830), ruthenium red (#R2751), RN1734 (#0658), SKF96365 (#S7809) and Pyr3 (#P0032) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The primary antibodies anti-TRPC1 (sc-133076), anti-TRPC3 (sc-514670), anti-TRPC4 (sc-15063), anti-TRPC5 (sc-18737), anti-Bax (sc-6236), and anti-Bcl-2 (sc-7382) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-TRPC6 (BA3394), anti-TRPC7 (PB0272) were purchased from Boster Biotechnology (Wuhan, China). Anti-caspase-3 (#9662), anti-Akt (#9272) and anti-pAkt(S473) (#4060) were from Cell Signaling Technology (Danvers, MA, USA). S-MEM without Ca2+ medium (#11380037) were obtained from Gibco. Other reagents were described as in specified.
7
Cholesterol and Lipid Micelle Delivery
8
Rat Cell Culture Reagents and Antibodies
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Sprague–Dawley rats were purchased from Charles River Japan (Hino, Japan). Dulbecco's modified Eagle's medium (DMEM) and DMEM/F-12 without phenol red, fetal bovine serum (FBS), trypsin-ethylenediaminetetraacetic acid (EDTA), and penicillin-streptomycin were purchased from GIBCO BRL (Grand Island, NY, USA). Human plasminogen and bovine fibrinogen were obtained from Enzyme Research Laboratories, Inc. (Swamsea, UK). Human urokinase and bovine thrombin were purchased from Calbiochem (Darmstadt, Germany). Monoclonal antibody for α-smooth muscle actin was purchased from DAKO (Glostrup, Denmark). 2′,7′-dichlorofluorescin diacetate (DCF-DA) was obtained from Molecular Probes (Eugene, OR, USA) and was dissolved in dimethyl sulfoxide (DMSO). 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]-pyrimidin-3-yl]phenol (PHTPP) were purchased from Tocris Bioscience (Bristol, UK). LysoPC and all other chemicals were obtained from Sigma Chemical Co. (St. Louis, MO, USA). E2 was dissolved in ethanol (EtOH), whereas ICI 182,780, MPP, and PHTPP were dissolved in DMSO. All of the other chemicals were dissolved in water.
9
Lysopc Modulation of HUVEC Cells
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Human umbilical vein endothelial cells (HUVECs), a human endothelial cell lineage, were obtained from the American Type Culture Collection (Rockville, MD, USA) and maintained in DMEM containing 10% FBS and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO2 at 37 °C. LysoPC (Sigma Aldrich, MS, USA) was dissolved in PBS and preserved at – 20 °C. When the cells reached approximately 80% confluence, they were treated with LysoPC for 24 h. Cells in the GDF11 and inhibitor groups were pretreated with GDF11 (50 ng/mL) and 4-PBA (5 mM) for 1 h prior to exposure to 50 ug/mL LysoPC for another 24 h.
10
Lipidomic Analysis of Cellular Metabolites
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Metabolites LysoPC and SM from egg yolk and internal standards were obtained from Sigma-Aldrich, with the exception of cytidine-5′-diphosphate, which was purchased from MP Biomedicals Inc. (Illkirch, France) and myo-inositol-1-phosphate from Interchim (Montluc, France). LysoPC contains primarily palmitic and stearic acids. SM contains primarily palmitic acid. All the deuterated compounds were purchased from CDN Isotopes (Québec, Canada). RPMI 1640 medium and RPMI 1640 without Cho, Ser, methionine, inositol, and folic acid were obtained from Gibco.
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