Egg phosphatidyl choline (EPC; Avanti Polar Lipids), the nickel lipid 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-([N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (NTA; Avanti Polar Lipids, Alabaster, AL), and Oregon green 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (DHPE; Molecular Probes, Eugene, OR) were combined in chloroform and dried in a glass container under N2 gas. Bilayers were prepared in either a 99.6% EPC/0.4% EPC ratio or a 91% EPC/8.6% NTA/0.4% DHPE proportion for FimA2 attachment. They were then resuspended in vesicle buffer and vortexed for 10 s. The solution was then sonicated for upward of 5 min to generate small, unilamellar vesicles (SUVs). The SUVs were then added to Piranha-treated coverslips to coat them with a phospholipid bilayer in a custom-built imaging chamber (Quorum Technologies, Guelph, Canada). The phospholipid bilayer was maintained hydrated in ∼500 μl of ATP-free F-buffer.
Egg phosphatidylcholine epc
Manufactured by Avanti Polar Lipids
Sourced in United States
Egg phosphatidylcholine (EPC) is a natural phospholipid derived from egg yolks. It is a major component of cell membranes and plays a crucial role in maintaining the structural integrity and fluidity of these membranes.
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Lab products found in correlation
2 iminothiolane hydrochloride traut s reagent, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Holo human tf, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Dotap, by Avanti Polar Lipids (1 mentions)
Dodma, by Avanti Polar Lipids (1 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine n methoxy polyethylene glycol 2000 dspe peg2000, by Avanti Polar Lipids (1 mentions)
U87 cell line, by American Type Culture Collection (1 mentions)
Cholesterol, by Merck Group (1 mentions)
Capsaicin cap, by Merck Group (1 mentions)
Lmp agarose, by Thermo Fisher Scientific (1 mentions)
1 1 dioctadecyl 3 3 3 3 tetramethylindodicarbocyanine 4 chlorobenzenesulfonate salt did, by Biotium (1 mentions)
1 2 dioleoyl sn glycero 3 phosphoethanolamine n carboxyfluorescein cfpe, by Avanti Polar Lipids (1 mentions)
6 protocols using egg phosphatidylcholine epc
1
Preparation of Phospholipid Bilayers
2
Targeted Delivery of Anti-Survivin siRNA
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Materials. 2-Chlorotrityl chloride resins and fluorenylmethyloxycarbonyl-L-arginine (Fmoc-Arg) were obtained from JiEr Biochemical Co. (Shanghai, China). Stearic acid (StA; 98.5%), cholesterol, holo-human Tf, 2-iminothiolane hydrochloride (Traut's reagent) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Egg phosphatidylcholine (ePC) was purchased from Avanti Polar Lipids (Alabaster, AL, USA). Distearylphosphatidylethanolamine-polyethyleneglycol (M.W. 2,000)-maleimide (DSPE-PEG2000-Mal) was purchased from Shanghai Advanced Vehicle Technology Pharmaceutical Ltd. Co. (Shanghai, China). Fetal bovine serum (FBS), Dulbecco's modified Eagle's medium (DMEM) and 0.25% (w/v) trypsin were from HyClone (Logan, UT, USA). Anti-survivin siRNA, and 6-carboxyfluorescein (FAM)labeled siRNA were synthesized by Ribo Biochemistry (Guangzhou, China). 4',6-Diamidino-2-phenylindole (DAPI) was purchased from Invitrogen Molecular Probes (Eugene, OR, USA). All other reagents were commercially purchased in reagent-grade.
3
Nanoparticle-based Doxorubicin Delivery
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Egg phosphatidylcholine (ePC), DOTAP, DODMA, DDAB, transferrin, and 1,2-distearoyl-sn- glycero-3-phosphoethanolamine-N-(methoxy-(polyethylene glycol)-2000) (PEG2000-DSPE) were all obtained from Avanti Polar Lipids (Shanghai, China). OA-R8 was synthesized by Jill Biochemical Co., Ltd. (Shanghai, China). Cholesterol and DOX were from Sinopharm Chemical Reagent co. Ltd. (Shanghai, China). The U87 cell line was from the American Type Culture Collection (ATCC, Rockefeller, MD, USA). Balb/c Nude mice (male, 6–8 weeks old, 18.0–20.0 g) were obtained from Shanghai Slack Laboratory Animals Co., Ltd. (Shanghai, China).
4
Lipid Nanoparticle Synthesis and Characterization
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Egg phosphatidylcholine (EPC) and (DSPE-PEG (2000)) (1,2- distearoyl-sn-glycero-3-phosphoethanolamine (polyethylene glycol)- 2000) were obtained from Avanti Polar Lipids (Alabaster, AL, USA). Cholesterol was purchased from Sigma (St. Louis, MO, USA) and 5(S),6(R),15(R)-trihydroxy-7E,9E,11Z,13E-eicosatetraenoic acid (15-epi-LipoxinA4) was obtained from Cayman Chemicals (Ann Arbor, MI, USA).
5
Extraction and Characterization of Cyanobacterial EPS
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The exopolysaccharides (EPS) (2 MDa) were obtained by the blue-green cynobacteria Neocyanospira capsulata, a strain of which was present in the culture collection of the Department of Agriculture, Food, Environment and Forestry (Microbiology Section) of the University of Florence, following the methodology described by Vincenzini et al. (1990 (link)). Egg phosphatidylcholine (EPC) was purchased from Avanti Polar Lipids Inc., Alabaster, AL. Capsaicin (CAP), with Molecular Weight (MW) of 305.41 g/mol, was purchased by Merck (Darmstadt, Germany) as well as all the solvents used. Hydroxytyrosol (HT), with MW of 154.16 g/mol, was extracted from the free-oil olive pulp at the PHYTOLAB, University of Florence, following the procedure described by Romani et al. (2017 (link)).
6
Cholesterol-Anchored Functional Nanomaterials
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Egg phosphatidylcholine (EPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(carboxyfluorescein) (CFPE) were purchased from Avanti Polar Lipids, Inc. (Alabaster, USA). Cholesterol (CHO) and cysteine (Cys) were purchased from Chengdu Kelong Chemical Company (Chengdu, China). All the cholesterol anchored functional materials were synthesized by our laboratory according to a previously reported method17 , including the cholesterol anchored reduction-sensitive PEG (CHO-S-S-PEG5000), R8 modified PEG (CHO-PEG2000-R8) and non-cleavable PEG2000 (CHO-mPEG2000). Agarose LMP was purchased from Invitrogen (Carlsbad, CA). 1,1'-Dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine 4-chlorobenzenesulfonate salt (DID) was obtained from Biotium (Hayward, CA). Other chemicals and reagents were of analytical grade.
Male BALB/c mice weighing 20–25 g were purchased from Experiment Animal Center of Sichuan University (China). All the animal experiments adhered to the principles of care and use of laboratory animals and were approved by the Experiment Animal Administrative Committee of Sichuan University.
Male BALB/c mice weighing 20–25 g were purchased from Experiment Animal Center of Sichuan University (China). All the animal experiments adhered to the principles of care and use of laboratory animals and were approved by the Experiment Animal Administrative Committee of Sichuan University.
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