1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) > 99% was purchased from Avanti Polar Lipids (Alabaster, AL, USA), cholesterol (CHOL) from Sigma-Aldrich (Taufkirchen, DE, Germany), and DIBO Alexa Fluor 488 from Invitrogen (Waltham, MA, USA). Commercially available chemical reagents, purchased from Sigma-Aldrich (Taufkirchen, DE, Germany), Alfa Aesar (Ward Hill, MA, USA), TCI (Eschborn, Germany), and Acros Organics (Fair Lawn, NJ, USA), were used as received without further purification. All solvents were distilled before usage and dried when needed using standard procedures. The moisture-sensitive reactions were carried out under nitrogen atmosphere. Analytical thin-layer chromatography (TLC) was performed using silica gel pre-coated aluminum plates with a thickness of 0.2 mm. The compounds were visualized with a potassium permanganate stain solution (1.50 g of KMnO4, 10.0 g of K2CO3, 100 mg of NaOH, and 200 mL of H2O). Liquid column chromatography purification was performed with silica gel 60 (40–63 μm mesh) from Macherey–Nagel (Düren, DE).
Cholesterol chol
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Italy
Cholesterol (Chol) is a laboratory equipment used to measure the levels of cholesterol in the blood. It provides accurate and reliable results to aid in the assessment of an individual's cardiovascular health.
Automatically generated - may contain errors
Lab products found in correlation
1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine popc, by Avanti Polar Lipids (5 mentions)
Tween 80, by Merck Group (5 mentions)
Chloroform, by Merck Group (5 mentions)
Span 60, by Merck Group (4 mentions)
Tween 60, by Merck Group (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Methanol, by Merck Group (4 mentions)
Hplc grade methanol, by Tedia (3 mentions)
Cellulose dialysis membranes, by HiMedia (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Phosphate buffered saline (pbs), by Merck Group (3 mentions)
Whatman, by Cytiva (3 mentions)
1 2 distearoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (3 mentions)
1 2 dipalmitoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Dotap, by Avanti Polar Lipids (2 mentions)
Fetal calf serum (fcs), by Merck Group (2 mentions)
Spectrapor dialysis membrane, by Serva Electrophoresis (2 mentions)
Calcein, by Merck Group (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
101 protocols using cholesterol chol
1
Synthesis and Purification of Fluorescent Lipid Probes
2
Preparation of Lipid Nanoparticles for siRNA Delivery
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Lipids dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA or MC3) were obtained from Med Chem Express (Monmouth Junction, NJ, USA). 1,2-Dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (poly-ethylene glycol)-2000]-maleimide (DSPE-PEG-Mal) were obtained from Avanti Polar Lipids (Alabaster, AL, USA). 1,1′-Diooctadecyl-3,3,3′,3′-teramethyl–indocarbocyanine perchlorate (DiI) as a lipophilic tracker and Hoechst 33,342 as a nucleic acid stain were obtained from Molecular Probes (Leiden, The Netherlands). Cholesterol (Chol) and N-succinimidyl S-acetylthioacetate (SATA) were purchased from Sigma (St. Louis, MO, USA). EGFP-S1 positive control DsiRNA (siRNAGFP) was purchased from Integrated DNA Technologies Inc. (Coralville, IA, USA). Other siRNA were obtained from Qiagen (Venlo, The Netherlands). LipofectamineTM 2000 was obtained from Invitrogen (Breda, The Netherlands). The hybridoma for producing E1/6-aa2 monoclonal antibody (mouse IgG1 against human VCAM-1) was kindly given by Dr. M. Gimbrone from the Harvard Medical School (Boston, MA, USA).
3
Synthesis of PEG-Peptidyl-Lipids and Controls
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Full details for the syntheses of PEG2000-peptidyl-lipids 1 -4 and two PEG2000-lipid controls 5 and 6 are described elsewhere (Yingyuad et al, 2013 (link)). The cationic lipid N’,N’-dioctadecyl-N-4,8-diaza-10-aminodecanoyl-glycylamide (DODAG) 7 was prepared as described previously (Mevel et al, 2010 ). Dioleoyl-L-α-phosphatidylcholine (DOPC) 8 and cholesterol (Chol) 9 along with all other chemicals were purchased from Sigma-Aldrich, Lancaster or Merck Biosciences. Anti-luciferase siRNA was obtained from Qiagen. Silencer negative control siRNA was purchased from Applied Biosystems/Ambion. HLE was purchase from Sigma/Aldrich (UK), MMP-2 from Calbiochem (UK). Propidium iodide and SYBR® green II RNA gel stain were obtained from Molecular Probes (UK). SYBR® safe DNA gel stain and 10x Tris-borate-EDTA (TBE) electrophoresis buffer were purchased from Invitrogen.
4
Lipid-based Nanoparticle Formulation Protocols
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Egg Phosphatidylcholine (PC), CAS: 8002–43–5, 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DPPG), CAS: 67232–81–9, 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), CAS: 63–89–8, and Cholesterol (Chol), CAS: 57–88–5, were obtained from Sigma-Aldrich Company Ltd. (Poole, UK). 1,2-dioleoyl-sn-glycero-3-phsphoEthanolamine (DOPE), CAS: 4004–05–1, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), CAS: 144189–73–1, dimethyldioctadecylammonium bromide (DDA), CAS: 3700–67–2, and trehalose 6,6-dibehenate (TDB), CAS: 66758–35–8 were purchased from Avanti Polar Lipids, Inc., (Alabaster, AL), purity >99% (Table I). Ethanol, CAS: 64–17–5, and mEthanol, CAS: 67–56–1, were obtained from Fisher Scientific (Loughborough, UK). TRIS Ultra Pure, CAS: 77–86–1, was obtained from ICN Biomedicals, Inc., (Aurora, Ohio, US). Propofol (2,6-Bis(isopropyl)phenol), CAS: 2078–54–8, and ovalbumin (chicken egg), CAS: 9006–59–1, were obtained from Sigma-Aldrich Company Ltd., (Poole, UK). Ultrafiltration regenerated cellulose membranes (p\n: U2755–10AE) were obtained from Sigma-Aldrich Company Ltd., (Poole, UK) (10 kDa, pore size 0.22 µm), and Biomax polyethersulfone ultrafiltration membrane discs with 300 kDa cutoff, pore size 0.45 µm (p\n: PBMK06210) from Merck Milipore (Darmstadt, Germany).
5
Preparation of Fluorescent Liposomes for Biological Studies
6
Siglec-7 Targeted Liposomal Delivery
7
Formulation and Characterization of Lipid Nanoparticles
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Cholesterol (Chol) was obtained from Sigma (St. Louis MO, USA). 1-methyl-4-(cis-9-dioleyl) methyl-pyridinium-chloride (SAINT-C18) was obtained from Synvolux Therapeutics Inc. (Groningen, The Netherlands). 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethyleneglycol)-2000] (DSPE-PEG2000) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000]-maleimide (Mal-PEG2000 -DSPE) were obtained from Avanti Polar Lipids (Alabaster, AL, USA).
8
Liposome Preparation and Composition
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Large unilamellar vesicles (LUVs), with a diameter of approximately 100 nm, were obtained by extrusion of multilamellar vesicles (MLVs) as described elsewhere [41 (link)]. 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-phosphoinositol (POPI), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) were from Avanti Polar Lipids (Alabaster, AL, USA), while cholesterol (Chol) and ergosterol (Erg) were from Sigma. The LUVs studied were mostly zwitterionic (pure POPC, POPC:Chol 70:30, POPC:Erg 70:30, and C. albicans membrane-like mixture, i.e., POPC:POPE:POPS:POPI:Erg 59:21:3:4:13 [42 (link)]), mimicking membrane systems relevant for the study [32 (link),41 (link)]. Liposomes were extruded in the day of the measurement. HEPES buffer was used in all the measurements, except when otherwise indicated.
9
Liposome Formulation for Drug Delivery
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1,2-Distearoyl-sn-glycerol-3-phosphatidyl-choline [DSPC], and 1,2-Distearoyl-sn-glycerol-3-phosphatidyl-ethanolamine-N-[methoxy(polyethylene-glycol)-2000] [PEG2000] were purchased from Lipoid, Germany. Cholesterol (Chol) was purchased from Sigma-Aldrich (Darmstadt, Germany). The rac-2,3-dipalmitoyl-oxypropylarsonic acid [ARS] (C16), was synthesized as described in detail before [1 (link),24 (link),25 (link)]; TREG lipid was also synthesized using the method reported in detail before [18 (link),19 (link)]. Doxorubicin, hydrochloric salt (DOX) was purchased by Tocris Bioscience, UK. Fetal Calf Serum (FCS) was from Sigma (Darmstadt, Germany).
A bath sonicator (Branson) and a microtip-probe sonicator (Sonics and Materials, Leicestershire, UK) were used for liposome preparation. Protein concentrations were measured by Bradford microassay (1–10 µg protein/mL). The Cell Viability MTT assay was carried out with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent, which was purchased by Sigma-Aldrich. All other reagents and chemicals used were of analytical grade, and were purchased from Sigma-Aldrich. The osmolarity of solutions used for liposome formation was measured by a Roebling osmometer and adjusted to 300 mOsm with NaCl (if needed).
A bath sonicator (Branson) and a microtip-probe sonicator (Sonics and Materials, Leicestershire, UK) were used for liposome preparation. Protein concentrations were measured by Bradford microassay (1–10 µg protein/mL). The Cell Viability MTT assay was carried out with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent, which was purchased by Sigma-Aldrich. All other reagents and chemicals used were of analytical grade, and were purchased from Sigma-Aldrich. The osmolarity of solutions used for liposome formation was measured by a Roebling osmometer and adjusted to 300 mOsm with NaCl (if needed).
10
PC Lipid Characterization and Membrane Thermodynamics
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