1 palmitoyl 2 oleoyl sn glycero 3 phosphoethanolamine

Manufactured by Avanti Polar Lipids

Sourced in United States

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine is a phospholipid compound. It is a structural component of biological membranes.

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Lab products found in correlation

82 protocols using 1 palmitoyl 2 oleoyl sn glycero 3 phosphoethanolamine

Lipid Stock Preparation and Handling

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1-Palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC), sodium salt of 1-palmitoyl-2-oleoyl-sn-glycero-3phospho-l-serine (POPS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1-2-dioleoyl-snglycerol (DAG) and brain ammonium salt of l-α-phosphatidylinositol-4,5-bisphosphate (PIP 2 ) were purchased from Avanti Polar Lipids. The lipid stock solutions were stored in glass containers layered with argon at -80 °C. Lipid-containing solutions were handled using glass microliter syringes (Hamilton).

Stankunas E, & Köhler A. (2024). Docking a flexible basket onto the core of the nuclear pore complex. Nature cell biology.

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Lipid Vesicle Preparation and Characterization

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Lipid vesicles were generated by mixing chloroform solutions of cholesterol (700000P); Npalmitoyl-D-erythro-sphingosylphosphorylcholine (SM-860584P); D-galactosyl-ß-1,1' Npalmitoyl-D-erythro-sphingosine (Cer-860521P); 1-palmitoyl-2-oleoyl-sn-glycerol (DG-800815C); 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate (PA-840857P); 1-palmitoyl-2-oleoylsn-glycero-3-phospho-L-serine (PS-840034P); 1-palmitoyl-2-oleoyl-glycero-3phosphocholine (PC-850457P) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (PS-850757P) from Avanti Polar Lipids (Alabaster, Alabama, USA) in the desired proportions. Lipids were dried under a stream of nitrogen followed by exposure to high vacuum for 30 min. Dried phospholipids were resuspended in the corresponding buffer (25 mM HEPES, pH 7.4 and 150 mM NaCl) by vigorous vortexing and also using a bath sonicator to get all of the lipid into suspension. Then, the lipid suspension was subjected to eight rounds of freezing (liquid N2)-thawing (in the bath sonicator at 40 °C). The large unilamellar phospholipid vesicles of ∼100 nm diameter were prepared by extruding rehydrated phospholipid suspensions through two stacked 0.1 μm polycarbonate membranes (Avanti Polar Lipids).

Egea-Jiménez A.L., Audebert S., Castro-Cruz M., Borg J., David G., Camoin L., & Zimmermann P. (2020). PLD2-phosphatidic acid recruit ESCRT-I to late endosomes for exosome biogenesis.

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Reconstituted Lipid Membrane Assay

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4-(2-Hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES), MgCl₂, CaCl₂, KCl, and ATP disodium
salt were all purchased from Sigma and used as received. HEPES buffer
(5.0 mM HEPES, 0.5 mM CaCl₂, 1.0 mM MgCl₂, pH
= 7) was prepared by dissolving the appropriate amount of chemicals
in ultrapure water (Milli-Q ultrapure water purification system).
The phospholipids, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine
(POPE), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (POPS) were obtained from Avanti Polar Lipids (Alabaster,
AL) and were stored in a −20 °C freezer until use. Recombinant
Hsc70 (Stressgen) was used as received without further purification
and was kept at −80 °C.

Macazo F.C, & White R.J. (2014). Monitoring Charge Flux to Quantify Unusual Ligand-Induced Ion Channel Activity for Use in Biological Nanopore-Based Sensors. Analytical Chemistry, 86(11), 5519-5525.

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Preparation of ECM-Mimicking Lipid Vesicles

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LUVs, mimicking the membrane domain involved in epithelial cell–ECM adhesion (Márquez et al. 2008 (link)), were prepared in pH 7.4 buffer containing 10.0 mM HEPES, 50.0 mM KCl, 1.0 mM EDTA, and 3.0 mM NaN₃, as previously described (Su et al. 2019). The lipid composition was with a lipid molar ratio of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC): 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE): 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS): 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoinositol(POPI) : sphingomyelin (SM) = 40:25:15:10:10 (Avanti Polar Lipids). Note that POPC, POPE, and SM have neutral charge, while POPS and POPI are negatively charged. Therefore, the ECM-adhesion-imitating LUVs used here are about 25% anionic. The size of the LUVs was 400 nm.

Su J., Bapat R.A., Visakan G, & Moradian-Oldak J. (2022). Coemergence of the Amphipathic Helix on Ameloblastin With Mammalian Prismatic Enamel. Molecular Biology and Evolution, 39(11), msac205.

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Preparation of OLVs and LUVs

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Oligolamellar (non-extruded) (OLVs), and unilamellar (extruded) vesicles (LUVs) of 1-palmitoyl-2oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG) (Avanti Polar Lipids, Alabama, USA) in a proportion 3:1 (mol/mol) were prepared as described before. 23

Silva T., Claro B., Silva B.F.B., Vale N., Gomes P., Gomes M.S., Funari S.S., Teixeira J., Uhríková D, & Bastos M. (2018). Unravelling a Mechanism of Action for a Cecropin A-Melittin Hybrid Antimicrobial Peptide: The Induced Formation of Multilamellar Lipid Stacks. Langmuir : the ACS journal of surfaces and colloids, 34(5).

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Antimicrobial Peptide Purification and Characterization

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Magainin 2 (GIGKFLHSAKKFGKAFVGEIMNS-NH₂) and pleurocidin (GWGSFFKKAAHVGKHVGKAALTHYL-NH₂) were purchased from Pepceuticals Ltd (Nottingham, UK) as desalted grade. Further HPLC purification was performed using acetonitrile/water gradients using either a Waters Symmetry™ C8, 5 μm, 7.8 × 100 mm column or a Waters SymmetryPrep™ C8, 7 μm, 19 × 300 mm column. The lipids 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3- phospho-(1′-rac-glycerol) (DMPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG), 1-palmitoyl_d31-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG-d31) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) were obtained from Avanti Polar Lipids, Inc. (Alabaster, AL) and used without further purification. All other reagents were analytical grade or better.

Amos S.B., Vermeer L.S., Ferguson P.M., Kozlowska J., Davy M., Bui T.T., Drake A.F., Lorenz C.D, & Mason A.J. (2016). Antimicrobial Peptide Potency is Facilitated by Greater Conformational Flexibility when Binding to Gram-negative Bacterial Inner Membranes. Scientific Reports, 6, 37639.

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Reconstitution of SNARE Proteoliposomes

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All lipids were obtained from Avanti Polar Lipids Inc. For t-SNARE reconstitution, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1-palmitoyl-2-oleoyl-sn-glyc-ero-3-phosphoserine (POPS) and cholesterol were mixed in a molar ratio of 60:20:10:10. For v-SNARE reconstitution, POPC, POPE, POPS, cholesterol, (N-(7-nitro-2,1,3-benzoxadiazole-4-yl)-1,2-dipalmi-toylphosphatidylethanolamine (NBD-DPPE) and N-(Lissamine rhod-amine B sulfonyl)-1,2-dipalmitoylphosphatidylethanolamine (rhod-amine-DPPE) were mixed at a molar ratio of 60:17:10:10:1.5:1.5. SNARE proteoliposomes were prepared by detergent dilution and isolated on a Nycodenz density gradient flotation.^{13 (link)} Complete detergent removal was achieved by overnight dialysis of the samples in Novagen dialysis tubes against the reconstitution buffer (25 mM HEPES [pH 7.4], 100 mM KCl, 10% glycerol, and 1 mM DTT). To prepare sulforhodamine-loaded liposomes, t- or v-SNARE liposomes were reconstituted in the presence of 50 mM sulforhodamine B (Sigma). Free sulforhodamine B was removed by overnight dialysis followed by liposome flotation on a Nycodenz gradient. The protein: lipid ratio was at 1:200 for v-SNAREs and at 1:500 for t-SNARE liposomes.

Yu H., Rathore S.S., Shen C., Liu Y., Ouyang Y., Stowell M.H, & Shen J. (2015). Reconstituting Intracellular Vesicle Fusion Reactions: The Essential Role of Macromolecular Crowding. Journal of the American Chemical Society, 137(40), 12873-12883.

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Lipid-Detergent Protein Purification Protocol

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Nitrilotriacetic acid (Ni²⁺-NTA)-chelating Sepharose and Superdex 200 were obtained from GE Healthcare. N-dodecylphosphocholine (DPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC), 1,2-dioleoyl-sn-glycero- 3-phospho-l-serine (PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (PE), 1,2-dioleoyl-sn-glycero-3-phospho-(1′-myo-inositol-4′,5′-bisphosphate) (PI(4,5)P₂), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(biotinyl) (biotin-PE), 1,2-dipalmitoyl-sn-glycero-3-phospho-ethanolamine-N-(7-nitro- 2-1,3-benzoxadiazol-4-yl) (NBD-PE) and N-(lissamine rhodamine B sulfonyl)-1,2- dipalmitoyl-sn-glycero-3-phosphoethanolamine (rhodamine-PE) were obtained from Avanti Polar Lipids. 2-Sulfonatoethyl methanethiosulfonate sodium salt (MTSES) was purchased from Biotium. All other chemicals were from Sigma.

Bao H., Goldschen-Ohm M., Jeggle P., Chanda B., Edwardson J.M, & Chapman E.R. (2015). Exocytotic fusion pores are composed of both lipids and proteins. Nature structural & molecular biology, 23(1), 67-73.

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Planar Lipid Bilayer Formation and CFTR Channel Reconstitution

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To prepare planar lipid bilayers, we drill a 0.2 mm hole in a Teflon cup and paint the hole with a phospholipid solution containing a 3 : 1 mixture of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (Avanti Polar Lipids) in n-decane. The lipid bilayer separates 1 ml of solution in the Teflon cup (cis side) from 5 ml of solution in the outer glass chamber (trans side). Both chambers are magnetically stirred and thermally insulated. We utilize a temperature control system (TC2BIP, Cell Micro Controls).
We transfer CFTR ion channels into the pre-formed lipid bilayer by spontaneous fusion of membrane vesicles containing the CFTR variants. To maintain uniform orientation and functional activity of the CFTR channels transferred into the bilayer, we add 2 mM ATP, 50 nM PKA, and membrane vesicles into the cis compartment only. We perform all measurements in symmetrical salt solution (300 mM Tris–HCl, pH 7.2, 3 mM MgCl₂ and 1 mM EGTA) under voltage-clamp conditions using an Axopatch 200B amplifier. We maintain a membrane voltage potential of –75 mV, the difference between cis and trans (ground) compartments. We analyze the resulting data as previously described.^{50 (link)}

Proctor E.A., Kota P., Aleksandrov A.A., He L., Riordan J.R, & Dokholyan N.V. (2014). Rational coupled dynamics network manipulation rescues disease-relevant mutant cystic fibrosis transmembrane conductance regulator †Electronic supplementary information (ESI) available. See DOI: 10.1039/c4sc01320d Click here for additional data file.. Chemical Science, 6(2), 1237-1246.

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Lipid Composition Analysis Protocol

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1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (phosphatidylethanolamine, PE), 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (phosphatidylcholine, PC), 1,2-dipalmitoyl-sn-glycero-3-phospho-ethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE), and N-(lissamine rhodamine B sulfonyl)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (Rho-PE) were purchased from Avanti Polar Lipids (Alabaster, AL).

Evans C.S., He Z., Bai H., Lou X., Jeggle P., Sutton R.B., Edwardson J.M, & Chapman E.R. (2016). Functional analysis of the interface between the tandem C2 domains of synaptotagmin-1. Molecular Biology of the Cell, 27(6), 979-989.

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