1 2 dioleoyl sn glycero 3 phosphocholine dopc

Manufactured by Merck Group

Sourced in United States

1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) is a synthetic lipid compound commonly used in scientific research and laboratory applications. It is a phospholipid that consists of two oleic acid chains attached to a glycerol backbone and a phosphocholine head group. DOPC is a versatile material that can be used to create model membrane systems, such as liposomes and lipid bilayers, for the study of various biological processes and membrane-related phenomena.

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6 protocols using 1 2 dioleoyl sn glycero 3 phosphocholine dopc

Fabrication of Lipid Bilayer Substrates

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1,2-dipalmitoyl-sn-glycero-3-phosphocoline semisynthetic (≥99%, DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (≥99%, DSPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (≥99%, DMPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lyophilized powder were acquired from Sigma-Aldrich Company (St. Louise, MO, USA) and used without further purification. Chloroform (99.0–99.4%), hydrogen peroxide (30–32%), sulfuric acid (95–97%, Emparta^®ACS) and water LiChrosolv^® for chromatography were obtained from Merck KGaA (Darmstadt, Germany).
Immersion oil for microscopy (UN 3082-9, PG III) was purchased from Merck Millipore (Billerica, MA, USA). Uncoated N-BK7 right angle prism (15 mm) was acquired from Edmund Optics (Barrington, NY, USA). Gold targets of 99.99% purity (Ø 57 mm × 0.2 mm) for argon sputter deposition was obtained from Ted Pella products (Redding, CA, USA).
A p-type <100> orientation silicon wafer was acquired from Siegert Wafer GmbH (Aachen, Germany). Substrate characteristics are: resistivity 10–20 Ω cm, thickness 525 ± 20 µm, diameter 100 ± 0.3 mm, prime grade, CZ growth (Czochralski process), boron doping (P/B type), Single Side Polishing (SSP) with 2 Semi-standard flats, Total Thickness Variation (TTV) <5 µm, bow <30 µm, warp <30 µm, particles < 10 of 0.3 µm.

González-Henríquez C.M., Villegas-Opazo V.A., Sagredo-Oyarce D.H., Sarabia-Vallejos M.A, & Terraza C.A. (2017). Thermal Response Analysis of Phospholipid Bilayers Using Ellipsometric Techniques. Biosensors, 7(3), 34.

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Quantifying Extracellular Vesicle Protein and Lipid

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We measured the amount of total protein in aliquots of EVs containing 5, 10, 20 and 50 × 10⁹ EVs using the BCA protein assay kit (Thermo Fisher Scientific) and determined EV numbers per microgram of protein. The total lipid content in EV preparations was measured by the modified sulpho-phospho vanillin (SPV) method, as described elsewhere [27 (link)]. Briefly, both standard (1,2-Dioleoyl-sn-glycero-3-phosphocholine, DOPC; Sigma, St. Louis, MO, USA) and hiPSC-NSC-EV samples in SEC buffer (40 μl each) were sonicated at 35 amplitude (20 kHz) for 10 minutes, intensely vortexed for 2 minutes and mixed with 200 μl of 96% sulphuric acid. Following brief vortexing, the tubes were incubated at 90°C for 20 minutes, moved to 4°C for 5 minutes and mixed with 120 μl of phospho-vanillin reagent and vortexed. Next, 280 μl of each sample was transferred to a 96 well plate, and the colour reaction was allowed to progress for 10 minutes at room temperature. Absorbance at 540 nm was determined with a plate reader. The total lipid content in EV samples was calculated using the standard graph, and these values were used to determine the protein-lipid ratio in different samples.

Upadhya R., Madhu L.N., Attaluri S., Gitaí D.L., Pinson M.R., Kodali M., Shetty G., Zanirati G., Kumar S., Shuai B., Weintraub S.T, & Shetty A.K. (2020). Extracellular vesicles from human iPSC-derived neural stem cells: miRNA and protein signatures, and anti-inflammatory and neurogenic properties. Journal of Extracellular Vesicles, 9(1), 1809064.

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Lipid Extraction and Purification

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1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), OA, and EA were bought from Sigma–Aldrich, Inc. (Gillingham, United Kingdom). The lipids had a purity of >99% and were used without further purification.

Tyler A.I., Greenfield J.L., Seddon J.M., Brooks N.J, & Purushothaman S. (2019). Coupling Phase Behavior of Fatty Acid Containing Membranes to Membrane Bio-Mechanics. Frontiers in Cell and Developmental Biology, 7, 187.

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Nanoliposome Adjuvant Synthesis Protocol

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MP adjuvant and MPQ adjuvant were prepared by the thin-film hydration method. To prepare MP adjuvant platform, 0.2 mg of monophosphoryl lipid A (MPL) (Avanti Polar Lipid, Alabaster, AL, USA), 1 mg of 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) (Sigma Aldrich, St. Louis, MO, USA), and 0.25 mg of cholesterol (Sigma Aldrich), were dissolved in 1 ml of ethanol. To prepare MPQ adjuvant platform, 0.2 mg of QS21 (Desert King International, San Diego, CA, USA) was added into the MP adjuvant platform. Then, the organic solvent was removed with a rotary evaporator from both adjuvant platform solutions to generate the thin film layer. The thin film was hydrated with 2 ml of PBS (Hyclone, Logan, UT, USA) at 60°C for 30 min, and the resulting solution was sonicated for 1 min with tip sonication under ice bath conditions. The resultant was rotated by tube revolver for 2 h. Finally, it was mixed with 2 mg of poly I:C (Invivogen, San Diego, CA, USA) dissolved in PBS at a 1:1 volume ratio, leading to the generation of MP or MPQ nanoliposome adjuvant.

Kwon K.W., Kang T.G., Lee A., Jin S.M., Lim Y.T., Shin S.J, & Ha S.J. (2023). Protective Efficacy and Immunogenicity of Rv0351/Rv3628 Subunit Vaccine Formulated in Different Adjuvants Against Mycobacterium tuberculosis Infection. Immune Network, 23(2), e16.

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Raman Imaging of Cartilage Composition

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Raman images (∼1000
× 400 μm (spatial resolution
of ∼2 μm)) of native articular cartilage and tissue engineered
constructs were measured by continuous scanning. Each Raman spectrum
was collected with an acquisition time of 0.3–0.5 s, 1 accumulation,
and a power on the sample of ∼41 mW using the 532 nm laser
excitation. No sample degradation was noticed using this power density.
For comparison, reference Raman spectra were also measured of chondroitin
sulfate (Sigma-Aldrich), collagen type II (Sigma-Aldrich), synthetic
hydroxyapatite (Sigma-Aldrich), 1,2-dioleoyl-sn-glycero-3-phosphocholine
(DOPC) (Sigma-Aldrich), glycogen (Sigma-Aldrich), and demineralized
H₂O using similar experimental parameters.

Bergholt M.S., St-Pierre J.P., Offeddu G.S., Parmar P.A., Albro M.B., Puetzer J.L., Oyen M.L, & Stevens M.M. (2016). Raman Spectroscopy Reveals New Insights into the Zonal Organization of Native and Tissue-Engineered Articular Cartilage. ACS Central Science, 2(12), 885-895.

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B16-F10 Melanoma Cell Line Protocol

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B16-F10 mouse melanoma cell line (ATCC^® CRL6475™) was purchased from the American Type Culture Collection (Manassas, VA, USA). Cytochalasin D, chlorpromazine hydrochloride, genistein, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DOPG), and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were purchased from Sigma (St. Louis, MO, USA). Mouse monoclonal anti-glycogen synthase kinase-3β (GSK-3β) (ab93926, epitope within amino acids 292–420); and rabbit monoclonal anti-caspase-3 (ab184787, epitope within amino acids 1–175 recognizing total caspase-3) were purchased from Abcam (Cambridge, MA, USA). In addition, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) was purchased from Avanti Polar Lipids (Alabaster, AL, USA). Ibidi^TM µ-Dish 35 mm, high ibiditreat were purchased from Ibidi (Fitchburg, WI, USA). Hoechst 33342, modified Harris Hematoxylin (72711), and eosin (71304) were purchased from Thermo Scientific (Rockford, IL, USA). Rodent Block M, Mouse-on-Mouse HRP-Polymer, Background Sniper, and Rabbit-on-Rodent HRP Polymer were purchased from Biocare Medical, (Pacheco, CA, USA).

Castañeda-Reyes E.D., Perea-Flores M.D., Dávila-Ortiz G, & Gonzalez de Mejia E. (2022). Liposomes Loaded with Amaranth Unsaponifiable Matter and Soybean Lunasin Prevented Melanoma Tumor Development Overexpressing Caspase-3 in an In Vivo Model. Pharmaceutics, 14(10), 2214.

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