Egg yolk phosphatidylcholine (PC), phosphatidylglycerol (PG), cholesterol (Chol), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine and paclitaxel were purchased from Sigma (St-Quentin-Fallavier, France). 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (Mal-PEG) and DOTAP were purchased from COGER (Paris, France). Docetaxel was purchased from VWR (Fontenay-sous-Bois, France). DIR, a fluorophore tag, was purchased from Caliper (Villebon-sur-Yvette, France). 2-Iminothiolane (Traut’s reagent) and Draq5 were purchased from Fisher Scientific (Illkirch-Graffenstaden, France). QuantiBRITE phycoerythrin (PE) and PE Mouse Anti-Human HER-2/neu were purchased from BD Biosci-ences (San Jose, CA, USA). Trastuzumab (Herceptin) was kindly given by Genentech (South San Francisco, CA, USA). All other reagents were of analytical grade.
Phosphatidylglycerol
Manufactured by Merck Group
Sourced in United States
Phosphatidylglycerol is a type of phospholipid found in biological membranes. It is a core component of the cell membrane and plays a role in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Phosphatidylethanolamine, by Merck Group (4 mentions)
Phosphatidylserine, by Merck Group (3 mentions)
Phosphatidylcholine, by Merck Group (3 mentions)
Egg yolk phosphatidylcholine, by Merck Group (2 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine, by Merck Group (2 mentions)
Docetaxel, by Avantor (2 mentions)
Paclitaxel, by Merck Group (2 mentions)
Cardiolipin, by Merck Group (2 mentions)
Phosphatidic acid, by Merck Group (2 mentions)
2 iminothiolane traut s reagent, by Thermo Fisher Scientific (2 mentions)
Phosphatidylinositol, by Merck Group (2 mentions)
B27 supplement, by Thermo Fisher Scientific (2 mentions)
Trastuzumab herceptin, by Genentech (1 mentions)
Draq5, by Thermo Fisher Scientific (1 mentions)
Quantibrite phycoerythrin pe, by BD (1 mentions)
Pe mouse anti human her 2 neu, by BD (1 mentions)
Oleic acid, by Merck Group (1 mentions)
Human tf iron free, by Merck Group (1 mentions)
N hydroxysuccinimide nhs, by Merck Group (1 mentions)
2 3 dioleoyloxy propyl trimethylammonium dotap, by Avanti Polar Lipids (1 mentions)
8 protocols using phosphatidylglycerol
1
Liposomal Formulation for Targeted Cancer Therapy
2
Preparation and Culture of HL60 Cells
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Drn, Lut, oleic acid (OA), phosphatidylglycerol, human Tf (iron-free), and N-hydroxysuccinimide (NHS) were purchased from Sigma (St Louis, MO, USA) and (2,3-dioleoyloxy-propyl)-trimethylammonium (DOTAP) was provided by Avanti Polar Lipids (Birmingham, AL, USA). PEG-COOH (NH2-PEG-COOH) and DSPE-PEG-COOH were obtained from Ponsure Biological (Shanghai, China). The human leukocyte cell line HL60 was obtained from the American Type Culture Collection (Manassas, VA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS at 37°C in a 5% CO2 incubator.
3
Liposomal Paclitaxel and Docetaxel Formulations
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Egg yolk phosphatidylcholine (PC), phosphatidylglycerol (PG), cholesterol (Chol), 1,2-distearoyl-sn-glycero- 3-phosphoethanolamine, and paclitaxel were purchased from Sigma-Aldrich Co (St Louis, MO, USA). 1,2-Distearoyl- sn-glycero-3-phosphoethanolamine-N-[maleimide (polyethylene glycol)]-2000 (Mal-PEG) was purchased from Coger (Paris, France). Docetaxel was purchased from VWR International (Fontenay-sous-Bois, France). DiR and DiL fluorophore tags were purchased from PerkinElmer Inc and Thermo Fisher Scientific (Waltham, MA, USA). 2-Iminothiolane (Traut’s reagent) was also purchased from Thermo Fisher Scientific. Trastuzumab was kindly provided by Genentech (South San Francisco, CA, USA). All other reagents were of analytical grade.
4
Cell Culture Reagents and Lipid Supplements
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Dulbecco's modified Eagle's medium (DMEM), penicillin/streptomycin, D-glucose, foetal bovine serum (FBS), L-glutamine, Trypsin, DNase I, corticosterone (CORT), Poly-L-Lysine, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), fibroblast growth factor (FGF), epidermal growth factor (EGF), and dimethyl sulfoxide were purchased from Sigma. Phosphatidylcholine (PC; from egg yolk), lysoPhosphatidylcholine (LPC; from egg yolk), phosphatidylserine (PS; from bovine brain), phosphatidylethanolamine (PE; from egg yolk), phosphatidylinositol (PI; from soy bean), phosphatidylglycerol (PG; from egg yolk), phosphatidic acid (PA; from egg yolk), sphingomyelin (SM; from egg yolk) and cardiolipin (CL; from bovine heart) were obtained in their respective salts from Sigma. B-27 supplement was obtained from Thermo Fisher Scientific.
5
Lipidomic Analysis of Membrane Extracts
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Identification of extracted membrane lipids and LPS was performed using a quadrupole time-of-flight tandem mass spectrometer (Q-TOF Maxis, Bruker Daltonics, Germany) with an updated collision cell for electrospray ionization source. Metabolite identification was confirmed by the fragmentation of the detected parent ions. External phospholipid standards (choline, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, and tetracycline; Sigma Aldrich) were used to optimize the MS/MS-acquisition conditions. We used the following databases for lipid identification: LIPID MAPS database (LIPID MAPS Lipidomics Gateway, a free resource sponsored by the National Institute of General Medical Sciences, USA; http://www.lipidmaps.org ), Byrdwell G. Resources for Lipid Analysis in the twenty-first Century (http://www.byrdwell.com ), Metlin (Scrips Center for Mass Spectrometry, USA; https://masspec.scripps.edu ) and Galactosylceramides and Glucosylceramides (Cerebrosides) (http://www.lipidhome.co.uk/ ).
6
Reconstitution of Reaction Centers in Liposomes
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RC-containing vesicles (proteoliposomes) were prepared according to Giustini et al. (2005) (link). Depending on the desired lipid in the sample, pure phosphatidylcholine (PC, 4.1 mg), phosphatidylglycerol (PG, 4.0 mg) or cardiolipin (CL, 4.0 mg) (all of maximum purity grade from Sigma) were dissolved in chloroform and dried under N 2 flow. Then the lipid film was resuspended in 5 mM KCl, 5 mM phosphate pH 7.0 buffer and sodium cholate 4% solution in the same buffer enough to reach a minimum ∼5 cholate/phospholipid molar ratio (in order to obtain uniform monodispersed mixed lipid-detergent micelles). After short sonication until clarity, RCs (dispersed in 10 mM TRIS, 100 mM NaCl, 0.01% DDAO, pH 8.0) were added to the suspension. Subsequent detergent removal by elution through a Sephadex G-50 (Pharmacia) column with 5 mM KCl, 5 mM phosphate pH 7.0 buffer led to RC-containing small unilamellar vesicles. Both the presence and concentration of RCs in the liposomes were checked by measurement of the 802 nm absorption band: in every experiment the RC concentration was constant in the (2.2 ± 0.2) μM range. Finally, the amount of UQ-10 necessary to reconstitute the Q B site at a minimum 0.90 occupancy fraction was added as aqueous solution in the presence of 30% TX-100, always checking for the slow phase fraction in the analysis of charge recombination measurements.
7
HPTLC Lipid Profiling Protocol
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Analytical grade organic solvents used in this study along with HPTLC chromatography plates (20 × 10 and 10 × 10 cm silica gel 60 F254 glass plates) were obtained from Merck (Darmstadt, Germany). Copper sulfate heptahydrate, potassium chloride, orthophosphoric acid (85%, w/w), and sulfuric acid (96% w/w) were obtained from CarlRoth (Karlsruhe, Germany). Lipid standards, namely, phosphatidylcholine (PC 15:0–18:1), phosphatidylethanolamine (PE 15:0–18:1), phosphatidylinositol (PI 18:1–18:1), phosphatidylserine (PS 18:1–18:1), phosphatidylglycerol (PG 16:0–18:1), diacylglyceryl trimethyl-homoserin (DGTS 16:0–16:0), monogalactosyldiacylglycerol (MGDG 18:1–18:1), digalactosyldiacylglycerol (DGDG 18:3–18:3 as main species), sulfoquinovosyldiacylglycerol (SQDG 18:3–16:0 as main species), eicosapentaenoic acid as a free fatty acid (FA), triolein as a triacylglycerol (TAG), di-oleic acid as a diacylglycerol (DAG), and monooleic acid as a monoacylglycerol (MAG), were purchased from Merck. The lipid standards were dissolved in n-hexane (p.a. ≥ 99%) and sonicated for 30 s. Stock solutions were prepared at a concentration of 1 mg mL−1 and aliquoted into 1.5-mL vials under a nitrogen stream, sealed, and stored at − 85 °C until analysis.
8
Quantitative Lipid Profiling by UHPLC-MS
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The lipids were determined according to our method described earlier [25 (link)] using an ExionLC AC (Sciex, Framingham, MA, USA) UHPLC system and a 4500 Q-TRAP mass spectrometer equipped with an ESI source. A total of 10 µL of lipid extract was injected onto an Eclipse XDB-C18, 50 mm × 4.6 mm, 1.8 µm column (Agilent, Santa Clara, CA, USA), heated at 40 °C, with the flow rate of 500 μL min−1. Ultrapure water (A) and methanol (B) were applied as a mobile phase, with both containing 5 mM ammonium formate. The solvent gradient was initiated at 70% B and, after 0.25 min, increased to 100% B for 1 min; then, it was maintained at 100% B for 6 min before being returning to the initial solvent composition over 2 min. The data analysis was conducted with Analyst v1.6.3 software (Sciex, Framingham, MA, USA). The phospholipids were determined qualitatively using an MRM method with the following standards for each class: phosphatidylethanolamine (PE 14:0/14:0; Merck, Darmstadt, Germany) and phosphatidylglycerol (PG 14:0/14:0; Merck, Darmstadt, Germany).
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