1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) (16:0 biotinyl-Cap-PE) lipids were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Lipids with a desired composition were mixed in chloroform, and subsequently dried in a stream of nitrogen gas. The mixed lipids were left overnight in a desiccator and were then subsequently hydrated with 2 mL of DI water. Small lipid vesicles, ~100 nm in diameter, were made by repeated freeze (liquid N2)/thaw (42°C) cycles followed by probe-sonication (9.9 s pulse; 3.3 s interval for 10 cycles at 30% max amplitude) in an ice bath, and then centrifuged at 20000 x g for 2 hr. ~1 mL of supernatant solution of small lipid vesicles was collected in fresh tubes and stored at 4°C after layering the tubes with argon gas.
1 2 dipalmitoyl sn glycero 3 phosphoethanolamine n cap biotinyl 16 0 biotinyl cap pe
Manufactured by Avanti Polar Lipids
Sourced in United States
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) (16:0 biotinyl-Cap-PE) is a phospholipid compound. It contains a biotin moiety coupled to the phosphoethanolamine head group through a spacer.
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3 protocols using 1 2 dipalmitoyl sn glycero 3 phosphoethanolamine n cap biotinyl 16 0 biotinyl cap pe
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Preparation of Biotinylated Lipid Vesicles
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Preparation of Biotinylated Lipid Vesicles
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Supported Lipid Bilayers for Cell Adhesion
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Methodologies of supported lipid bilayer preparation and membrane functionalization have been described in Yu et al. (2011) (link) and Yu et al. (2013) (link). In brief, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) 16:0 (biotinyl-Cap-PE) were purchased from Avanti Polar Lipids, Inc. The lipids (0.2 mol% biotinyl-Cap-PE and 99.8 mol% DOPC) were mixed with an equal volume of PBS and pipetted onto cleaned glass substratum with a 25-mm coverslip placed on top for self-assembly of lipid vesicles. The lipid-coated coverslips were immersed into a deionized water bath and then placed and sealed in an Attofluor cell chamber (Thermo Fisher Scientific). The supported lipid bilayer membrane ensemble was kept under aqueous environment at all times. For membrane functionalization, the supported lipid membrane was blocked with 50 µg/ml Casein. A total of 0.1 µg/ml Cascade blue neutravidin (Thermo Fisher Scientific) was added onto supported lipid membranes, followed by 1 µg/ml biotinylated RGD, cyclo (Arg-Gly-Asp-d -Phe-Lys [Biotin-PEG-PEG]) (Peptides International). Cells were then added onto the RGD-functionalized lipid bilayer membrane and imaged or fixed within 2–3 h of preparation.
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