Palmitoyloleoylphosphatidylcholine (POPC) (Avanti Polar Lipids Inc., Alabama, USA) - Lipid vesicles were produced according to Huppa et al. [30 (link)] and kindly provided to us by the Institute of Applied Physics at the Vienna University of Technology. After applying 100 μl phosphate buffered saline (PBS) buffer to the chamber of the lipid slides, 10 μl lipid is added for 15 minutes allowing the lipid to coat the surface. Then, the sample is rinsed with buffer to remove free lipid vesicles.
Palmitoyl oleoyl phosphatidylcholine popc
Manufactured by Avanti Polar Lipids
Sourced in United States
Palmitoyl-oleoyl-phosphatidylcholine (POPC) is a phospholipid molecule consisting of a glycerol backbone, a phosphocholine head group, and two fatty acid chains (palmitoyl and oleoyl). This lipid is commonly used in research to model cell membranes and study membrane-related biological processes.
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Lab products found in correlation
Palmitoyl oleoyl phosphatidylglycerol popg, by Avanti Polar Lipids (3 mentions)
Sodium cholate, by Merck Group (2 mentions)
Fructose cas no 57 48 7, by Merck Group (2 mentions)
N ε carboxymethyllysine cas no 941689 36 7, by Merck Group (2 mentions)
Lipoprotein lipase lpl from porcine pancreas, by Merck Group (2 mentions)
Quercetin 4 o glucoside, by Merck Group (1 mentions)
Quercetin, by Extrasynthese (1 mentions)
Cyanidin 3 o glucoside, by Phytolab (1 mentions)
Zebron zb 5 column, by Phenomenex (1 mentions)
8 aminonaphthalene 1 3 6 trisulfonic acid, by Thermo Fisher Scientific (1 mentions)
Neb q5 site directed mutagenesis kit, by New England Biolabs (1 mentions)
Nanodrop onec, by Thermo Fisher Scientific (1 mentions)
Phospholipase a2 from bee venom, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by ITW Reagents (1 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
Dimyristoylphosphatidylcholine, by Avanti Polar Lipids (1 mentions)
Lowry assay, by Merck Group (1 mentions)
Peptide synthesis reagents, by Thermo Fisher Scientific (1 mentions)
Cuso4, by Merck Group (1 mentions)
2 phenoxyethanol, by Merck Group (1 mentions)
12 protocols using palmitoyl oleoyl phosphatidylcholine popc
1
POPC Lipid Vesicle Formation
2
Lipid Bilayer Preparation Protocol
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Dioleoyl-phosphatidylserine (DOPS), dioleoyl-phosphatidylethanolamine (DOPE), salts, and other basic chemicals used in PLM studies were obtained from Sigma (Munich, FRG, analytical grade); palmitoyl-oleoyl-phosphatidylcholine (POPC) was purchased from Avanti Polar Lipids, (Alabaster, AL, USA). Quercetin was bought from Extrasynthese, Quercetin-4′-O-glucoside from Sigma-Aldrich, and cyanidin and cyanidin-3-O-glucoside were bought from Phytolab GmbH & Co.KG (Vestenbergsgreuth, Germany). All other chemicals were of high purity from commercial sources.
3
Characterization of Policosanol Compounds
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Palmitoyloleoyl phosphatidylcholine (POPC, #850457) was supplied by Avanti Polar Lipids (Alabaster, AL, USA). Sodium cholate (#C1254) was procured from Sigma (St Louis, MO, USA). N-ε-carboxymethyllysine (CAS-No 941689-36-7, Cat#14580-5g) and fructose (CAS-No 57-48-7, Cat #F0127) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Policosanol 1, sugar cane wax alcohol, was obtained from National Center for Scientific Research (CNIC), Habana, Cuba. Policosanols 2 and 3 were from sugar cane and supplied by Xi’an Natural Field Biotechnique Co., Ltd. (Xi’an, China) and Xi’an Realin Biotechnology Co., Ltd. (Xi’an, China), respectively. Policosanol 4, from rice bran, was purchased from Shaanxi Pioneer Biotech (Xi’an, China). All raw materials of each policosanol were analyzed using the same procedure by gas chromatography (HP-5890A GC, Agilent, Palo Alto, CA, USA) with a GC-flame ionization detector and a Zebron ZB-5 column (30 m × 0.53 mm × 1.50 μm) from Phenomenex (Torrance, CA, USA) at the Korea Advanced Food Research Institute (Uiwang-si, Republic of Korea). Certificates of analysis are available in the supplementary files .
4
Fluorescent Lipid Membrane Assay
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Phospholipids used in this study: palmitoyl-oleoyl-phosphatidylcholine (POPC) and palmitoyl-oleoyl-phosphatidylglycerol (POPG) were purchased from Avanti Polar Lipids (Alabaster, AL, USA). The fluorescent dye 8-aminonaphthalene-1,3,6-trisulfonic acid, disodium salt (ANTS), and quencher p-xylene-bis-pyridinium bromide (DPX) were obtained from Invitrogen (Carlsbad, CA, USA).
5
Fluorescence Studies of Protegrin Variants
6
Sortase A Pentamutant and MSP1D1ΔH5 Expression
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The construct for the evolved pentamutant of Sortase A in a pET29 vector was a gift from Prof. David R. Liu's laboratory (Harvard University). Expression and purification of the pentamutant Sortase A from Escherichia coli BL21(DE3) cells was performed as previously described (Chen et al., 2011 (link)). The construct for MSP1D1ΔH5 (dH5 hereafter) in a pET28a vector was a gift from Prof. Gerhard Wagner at the Harvard Medical School. Using NEB Q5 Site-Directed Mutagenesis Kit (New England BioLabs, NEB), a DNA sequence encoding LPGTGAAALEHHHHHH was appended into the end of the encoding sequence for MSP1D1ΔH5 construct to create the NW9 construct. Synthetic lipids, palmitoyloleoyl-phosphatidylcholine (POPC) and palmitoyloleoyl-phosphatidylglycerol (POPG), in powder form were purchased from Avanti Polar Lipids (Alabaster, AL). These were resuspended in chloroform and dried by nitrogen flow and vacuum overnight to a thin layer in glass tubes. Lipid stocks were sealed and left at −80°C for long-term storage.
7
Phospholipase A2 Enzyme Kinetics
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Palmitoyloleoylphosphatidylcholine (POPC) was purchased from Avanti Polar Lipids (USA); Phospholipase A2 from bee venom was obtained from Sigma-Aldrich. Enzyme stock solution (1.0 mM) was prepared in 0.1 M Tris-HCl buffer containing 0.1 M NaCl, pH 8.5. Enzyme concentration was controlled by the absorption at 280 nm (Nano Drop OneC, Thermo Fisher). 2-Amino-2-(hydroxymethyl)propane-1,3-diol (Tris base, Fluka), HCl, NaOH, ethylenediaminetetraacetic acid (EDTA, Panreac), and CaCl2 were of reagent grade.
8
Oxidative Modification of Lipoproteins
9
Preparing Lipid Samples with Cosolvents
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The phospholipids dimyristoylphosphatidylcholine (DMPC) and palmitoyl-oleoyl-phosphatidylcholine (POPC) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). Urea and TMAO were obtained from Sigma-Aldrich and NaCl from VWR. The samples were prepared so that the co-solute-lipid mass fraction (of urea, TMAO or NaCl) in the dry samples, C lip cosol = m cosol /(m cosol + m lip ), was fixed while the water content in terms of the number of water molecules per lipid n w = N w /N lip is variable. The samples were mixed together by the use of a mortar and pestle to produce a fine powder, which was then subjected to drying under vacuum with a 3 Å molecular sieve for at least 24 hours. According to earlier reports, 27, 28 this drying procedure removes all water from the lipid sample. The dry samples were then loaded into the measurement device under a stream of dry N 2 .
10
Peptide Synthesis and Surfactant Extraction Protocol
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