Neutral LUVs were prepared using 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC; Avanti Polar Lipids, Alabaster, AL, USA), whereas anionic ones were prepared using a 3:1 (mol:mol) mixture of POPC and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS; Avanti Polar Lipids, Alabaster, AL, USA) (Figure 1 ). Lipids were (co)dissolved in 3:7 (v:v) methanol:chloroform in a glass vial. Solvents were evaporated under a stream of nitrogen, after which the resulting lipid film was further dried under reduced pressure (0.8 bar) overnight. The lipids were then hydrated with 20 mM TRIS buffer at pH 7.4 to a total lipid concentration of 30 mM, and the sample was then subjected to five freeze–thaw cycles before being extruded 31 times through a 0.1 μm polycarbonate membrane filter (Avanti Polar Lipids, Alabaster, AL, USA).
1 palmitoyl 2 oleoyl sn glycero 3 phospho l serine pops
Manufactured by Avanti Polar Lipids
Sourced in United States
1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) is a phospholipid compound. It is a naturally occurring lipid found in cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine popc, by Avanti Polar Lipids (2 mentions)
1 palmitoyl 2 oleoyl glycero 3 phosphocholine popc, by Avanti Polar Lipids (1 mentions)
Paclitaxel, by Cayman Chemical (1 mentions)
Isopropyl β d 1 thiogalactopyranoside iptg, by GoldBio (1 mentions)
Gel extraction kit, by Qiagen (1 mentions)
Ni nta resin, by GoldBio (1 mentions)
E coli dh5α, by Thermo Fisher Scientific (1 mentions)
5 6 carboxyfluorescein cf, by Merck Group (1 mentions)
1 palmitoyl 2 oleoyl sn glycero 3 phosphatidylcholine popc, by Avanti Polar Lipids (1 mentions)
Nanosight lm14, by Malvern Panalytical (1 mentions)
1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
1 palmitoyl 2 oleoyl sn glycero 3 phosphoethanolamine, by Avanti Polar Lipids (1 mentions)
Polycarbonate membrane, by Cytiva (1 mentions)
5 protocols using 1 palmitoyl 2 oleoyl sn glycero 3 phospho l serine pops
1
Preparation of Neutral and Anionic Liposomes
2
Recombinant Human CYP2C8 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human CYP2C8 gene cloned into the AmpR pAr5 (modified pCWOri+) plasmid was a gift from Dr. Eric Johnson. PCR reagents were purchased from New England Biolabs (Ipswich, MA, USA). Molecular biology enzymes and E. coli DH5α were purchased from Invitrogen (Waltham, MA, USA). Plasmid DNA was purified using a Qiagen Gel Extraction kit. Ampicillin (Amp), arabinose, chloramphenicol (Chlr), isopropyl β-D-1-thiogalactopyranoside (IPTG), and Ni-NTA resin were purchased from Gold Biotechnology (St. Louis, MO, USA). δ-Aminolevulinic acid (δ-ALA) was purchased from Frontier Scientific (Emeryville, CA, USA). 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Carbamazepine and paclitaxel were purchased from Cayman Chemicals (Ann Arbor, MI, USA). NADPH was purchased from P212121 Store.
3
Liposome Preparation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liposomes were prepared according to methods that are well established in the field [17 , 18 (link)]. Briefly, liposomes were prepared by dry film formation, hydration and finally extrusion through a polycarbonate membrane to form monodisperse large unilamellar vesicles. The lipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) (Avanti Polar Lipids, Alabaster, USA) were mixed at molar ratios 1:99, 5:95 and 10:90 while dissolved in chloroform. A dry lipid film was formed by evaporation of the chloroform by nitrogen flow and overnight lyophilization. The film was hydrated with either 10 mM phosphate buffer (PB) pH 7 or 10 mM phosphate buffer saline (PBS) pH 7, and the solution was vortexed for 1 min and put on a shaker for 1 h before extruded 21 times through a 100 nm pore-sized polycarbonate membrane. For fluorescence leakage assay the lipid film was hydrated with buffer (PBS) containing self-quenching concentration (50 mM) of 5 (6)-carboxyfluorescein (CF) (Sigma Aldrich) and liposomes were prepared as described above. Removal of unencapsulated CF was done by gel filtration using a PD-25 column (GE Healthcare, Uppsala, Sweden) and liposomes with encapsulated CF were eluted with PBS.
4
Synthetic Liposome Preparation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Synthetic liposomes were prepared using a modified version of a previously described method52 . Briefly, chloroform suspended 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) (Avanti Polar Lipids, Alabaster, AL) were combined to form lipid mixtures of the described molar ratios. Lipid solutions were then dried to a thin film under a slow N2 flow and vacuum desiccated for 1 hour. Lipid films were resuspended in HBS buffer (10 mM HEPES, 0.15 M NaCl, pH 7.4) containing 2 mM CaCl2. Homogenously sized liposomes were formed by extruding the lipid solutions through polycarbonate track etched membranes (GE Healthcare, Pittsburgh, PA) with pore sizes of 30, 100, and 400 nm using a LiposoFast FL-50 extruder (Avestin, Ottawa, Canada). Extruded liposome sizes were characterized by nanoparticle tracking analyses using a NanoSight LM14 (Malvern Instruments, Malvern, United Kingdom).
5
Liposome-based EphA2 Kinase Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liposomes were prepared by drying 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) (Avanti Polar lipids), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) (Avanti Polar lipids), phosphatidylinositol 4,5-bisphosphate diC16 (PIP2) (Echelon Biosciences) and phosphatidylinositol 3,4,5-trisphosphate diC16 (PIP3) (Echelon Biosciences) in the desired ratios (w/w) overnight under vacuum. The lipid films were re-suspended in buffer (20mM HEPES, pH 7.4, 100mM NaCl) and subjected to 7 cycles of freeze-thaw using liquid nitrogen to generate liposomes. Liposomes were then extruded by passing them through a 0.1μm Polycarbonate membrane (Whatman). Final lipid concentrations were 2mg/ml. 100μl of liposomes were mixed with 50μl of EphA2 kinase domain protein (0.1mg/ml) and incubated at room temperature for 1 hour. Liposome-protein mixtures were centrifuged at 150,000×g for 30minutes at 20°C. Pellets were washed vigorously with buffer (20mM HEPES, pH 7.4, 100mM NaCl) and centrifuged again. These experiments were repeated 5 times and for each set the bound protein fractions was analysed by SDS-PAGE. The protein bands on gels were quantified by densitometry using Image Lab software. Averages of the ratio of intensities (bound: unbound) and SEM from all experiments were then calculated.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!