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1 2 dioleoyl sn glycero 3 phosphoethanolamine dope

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1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) is a synthetic phospholipid used in laboratory research and applications. DOPE is a neutral, zwitterionic lipid that can form non-lamellar, hexagonal phase structures. It is commonly used as a component in liposome and nanoparticle formulations.

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Lab products found in correlation

1 2 dioleoyl sn glycero 3 phosphocholine dopc, by Avanti Polar Lipids (8 mentions) Cholesterol, by Merck Group (7 mentions) Cholesterol, by Avanti Polar Lipids (5 mentions) 1 2 dioleoyl sn glycero 3 phospho 1 rac glycerol dopg, by Avanti Polar Lipids (3 mentions) C14 peg2000, by Avanti Polar Lipids (3 mentions) Milli q system, by Merck Group (3 mentions) Dmg peg 2000, by NOF corporation (3 mentions) Corn trypsin inhibitor, by Enzyme Research (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) 1 2 dimyristoyl sn glycerol methoxyethyleneglycol 2000 ether peg dmg, by NOF corporation (3 mentions) 100 nm polycarbonate membrane, by Cytiva (2 mentions) Chloroform, by Merck Group (2 mentions) 1 2 dimyristoyl sn glycero 3 phosphoethanolamine n methoxy polyethylene glycol 2000 peg, by Avanti Polar Lipids (2 mentions) Pump 33 ds syringe pumps, by Harvard Apparatus (2 mentions) 1 2 dioleoyl sn glycero 3 phospho l serine dops, by Avanti Polar Lipids (2 mentions) Pnp peg3400 pnp, by Laysan Bio (2 mentions) Hepes, by Merck Group (2 mentions) Xenolight d luciferin potassium salt, by PerkinElmer (2 mentions) 3 morpholinopropane 1 sulfonic acid mops, by Nacalai Tesque (2 mentions) Sodium chloride (nacl), by Merck Group (2 mentions)

46 protocols using 1 2 dioleoyl sn glycero 3 phosphoethanolamine dope

1

Preparation of Fluorescent Lipid Bilayer Vesicles

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A lipid film composed of 1,2‐dioleoyl‐sn‐glycero‐3‐phosphocholine (DOPC) (Avanti Polar Lipids), 1,2‐dioleoyl‐sn‐glycero‐3‐phosphoethanolamine (DOPE) (Avanti Polar Lipids) and cholesterol (Sigma Aldrich) with a mass ratio of 50:25:25 (total mass 1.25 mg) and 2 mol% additional 1,1′‐Dioctadecyl‐3,3,3′,3′‐Tetramethylindodicarbocyanine Perchlorate (DiD’) (Invitrogen, Thermo Fisher Scientific) was prepared via evaporation from a chloroform solution in a glass vial. After being thoroughly dried by lyophilisation (Labconco), it was rehydrated (1.25 mg/ml) in TE buffer (pH 8.0) containing 10 mM Tris‐HCl (Sigma‐Aldrich), 1 mM EDTA (Sigma‐Aldrich) and 150 mM NaCl (Sigma‐Aldrich) at 37 ˚C for 1 h. After being fully resuspended by vortex, the emulsion was extruded 31 times through a 100 nm polycarbonate membrane (Whatman) at 37 ˚C using an Avanti MiniExtruder (Avanti Polar Lipids). The liposome suspension was stored in a low adsorption glass vial (Supelco, Sigma‐Aldrich) at 4 ˚C protected from light, until use.
Görgens A., Corso G., Hagey D.W., Jawad Wiklander R., Gustafsson M.O., Felldin U., Lee Y., Bostancioglu R.B., Sork H., Liang X., Zheng W., Mohammad D.K., van de Wakker S.I., Vader P., Zickler A.M., Mamand D.R., Ma L., Holme M.N., Stevens M.M., Wiklander O.P, & EL Andaloussi S. (2022). Identification of storage conditions stabilizing extracellular vesicles preparations. Journal of Extracellular Vesicles, 11(6), e12238.
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2

Liposomal Formulation Development

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All the lipids, including 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (DSPE-PEG2000-MAL), a mini extruder, and polycarbonate membrane filters were purchased from Avanti Polar Lipids Inc. Cholesterol, protamine, and chloroform were purchased from Sigma-Aldrich. Fetal bovine serum (FBS), Dulbecco’s Modified Eagle Medium (DMEM), SYBR Safe, and lipofectamine 2000 were purchased from Invitrogen.
Wang D.D., Yang M., Zhu Y, & Mao C. (2015). Reiterated Targeting Peptides on the Nanoparticle Surface Significantly Promote Targeted Vascular Endothelial Growth Factor Gene Delivery to Stem Cells. Biomacromolecules, 16(12), 3897-3903.
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3

Formulation and Characterization of Lipid Nanoparticles

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Lipid nanoparticles (LNPs) were formulated by preparing an ethanol phase containing C14–4 ionizable lipid, 1,2-Dioleoyl-sn-glycero-3-phos-phoethanolamine (DOPE) (Avanti Polar Lipids, Alabaster, AL), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG) (Avanti Polar Lipids), cholesterol (Avanti Polar Lipids), and X-hydroxycholesterol. An aqueous phase was prepared containing 25 μg luciferase mRNA in 300 μL of 10 mM citric acid. LNPs were formulated via chaotic mixing of the ethanol and aqueous phases in a microfluidic device at a 1:3 volume ratio using pump33DS syringe pumps (Harvard Apparatus, Holliston, MA) [58 (link)]. LNPs were subsequently dialyzed against 1× PBS in 20 kDa molecular weight cutoff dialysis cassettes for 2 h and sterile filtered through 0.22 μm filters.
Patel S.K., Billingsley M.M., Frazee C., Han X., Swingle K.L., Qin J., Alameh M.G., Wang K., Weissman D, & Mitchell M.J. (2022). Hydroxycholesterol substitution in ionizable lipid nanoparticles for mRNA delivery to T cells. Journal of controlled release : official journal of the Controlled Release Society, 347, 521-532.
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4

Lipid Monolayer Characterization in PBS

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(DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine
(DOPE), and cholesterol were purchased from Avanti Polar Lipids. Chloroform,
methanol (both high-performance liquid chromatography (HPLC) grade),
D2O (99.9% deuterium content), buffer salts, and HCl and
DCl (99% deuterium content) for pH/pD adjustment were purchased from
Sigma-Aldrich. All water was ultrapure with resistance ≥ 18
MΩ and TOC ≤ 2 ppm produced from a Milli-Q Reference
A+ purification system. All monolayer and IR experiments were carried
out in phosphate buffered saline of the following composition: 150
mM NaCl, 20 mM PO43– in H2O (phosphate-buffered saline, PBS) or D2O (d-PBS) at pH/pD
7.4, respectively.
Daudey G.A., Schwieger C., Rabe M, & Kros A. (2019). Influence of Membrane–Fusogen Distance on the Secondary Structure of Fusogenic Coiled Coil Peptides. Langmuir, 35(16), 5501-5508.
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5

Preparation of Liposome Nanoparticles

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1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS) (Avanti Polar Lipids), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) (Anatrace), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) (Avanti Polar Lipids) and cholesterol (Sigma) were dissolved in chloroform and mixed (3:2:3:2 w/w ratio). The lipid mix was dried under nitrogen gas and rehydrated in Buffer A. Liposomes were extruded through a 100 nm polycarbonate membrane (Whatman) to create a monodisperse unilamellar liposome population.
Boyd C.M., Parsons E.S., Smith R.A., Seddon J.M., Ces O, & Bubeck D. (2016). Disentangling the roles of cholesterol and CD59 in intermedilysin pore formation. Scientific Reports, 6, 38446.
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6

Reconstitution of RyR2 with DP_cpvtN2

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1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (cis)—DOPC, 1,2-diphytanoyl-sn-glycero-3-phosphatidylcholine (DPPC), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were from Avanti Polar Lipids (USA).
The peptide DPcpvtN2, containing amino acids 410–438 of human RyR2 with the sequence HEESRTARVIRSTVFLFBRFIRGLDALSK was synthesized by GenScript (USA). Aliquots of a stock solution of the lyophilized peptide in water (100 μmol/l) were kept at −20°C.
Faltinova A., Tomaskova N., Antalik M., Sevcik J, & Zahradnikova A. (2017). The N-Terminal Region of the Ryanodine Receptor Affects Channel Activation. Frontiers in Physiology, 8, 443.
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7

Membrane Lipid Composition Analysis

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1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rh-DOPE), and 7-nitro-2-1,3-benzoxadiazol-4-yl (NBD)–DOPE were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Annexin V (Alexa Fluor 488-conjugated) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). α-Hemolysin from Staphylococcus aureus was purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA).
Kamiya K., Osaki T, & Takeuchi S. (2019). Formation of vesicles-in-a-vesicle with asymmetric lipid components using a pulsed-jet flow method. RSC Advances, 9(52), 30071-30075.
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8

Thrombin Generation Assay with TFPI Inhibition

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Thrombin calibrator standard was obtained from Thrombinoscope BV (Maastricht, the Netherlands). Fluorogenic substrate I-1140 was obtained from BACHEM (Bubendorf, Switzerland). TF was supplied by Dade Innovin® (Dade Behring, Marburg, Germany). Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoserine (DOPS), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were obtained from Avanti Polar Lipids (Delfzijl, The Netherlands). Phospholipid vesicles (20% DOPS, 20% DOPE, and 60% DOPC) were prepared as described. Corn trypsin inhibitor (CTI) was from Haematologic Technologies Inc (Vermont, USA). Monoclonal antibodies against TFPI were purchased from Sanquin (Amsterdam, the Netherlands) and pooled together to prepare an anti-TFPI antibody cocktail consisting of 4 monoclonal antibodies directed against different epitopes of TFPI (clone CLB/TFPI Kunitz-1, Kunitz-2, Kunitz-3 and C-terminus). Truncated TFPI (TFPI1-161) and full length TFPI were kind gifts from Dr. Lindhout and Prof. Dr. Buurman from the Faculty of Health, Medicine and Life Sciences, respectively. Convulxin was obtained from Enzo Life Sciences BVBA (Antwerp, Belgium). Hirudin was obtained from Nodia BV (Antwerp, Belgium).
Winckers K., Thomassen S., ten Cate H, & Hackeng T.M. (2017). Platelet full length TFPI-α in healthy volunteers is not affected by sex or hormonal use. PLoS ONE, 12(2), e0168273.
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9

Cationic Lipid-Peptide Nanoparticle Formulation

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1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DPPE-PEG2000) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Peptide ME27 (K16RVRRGACRGDCLG) was synthesized by Alta Bioscience (Birmingham, UK).
Meng J., Tagalakis A.D, & Hart S.L. (2020). Silencing E3 Ubiqutin ligase ITCH as a potential therapy to enhance chemotherapy efficacy in p53 mutant neuroblastoma cells. Scientific Reports, 10, 1046.
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10

Lipid Nanoparticle Preparation Protocol

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Commercial AgBh powder (Thermo Fisher Scientific) was used without any further purification. 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) was purchased from Avanti Polar Lipids Inc. The lipids were dissolved in water (DDW), using a total lipid concentration was 30 mg/ml per sample. The samples were homogenized using a vortexer for 5 minutes at 3000 RPM and were filled in quartz capillaries with a diamter of 1.5 mm, containing about 40μl .
Gutman B., Mrejen M., Shabat G., Avinery R., Shkolnisky Y, & Beck R. (2020). Angular super-resolution retrieval in small-angle X-ray scattering. Scientific Reports, 10, 16038.
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