Synthetic 1,2-dilauroyl-sn-glycero-3-phosphcocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dilauroyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DLPG), 1,2-dimyrsitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DMPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), 1,2-distearoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DSPG), 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG), 1,2-distearoyl-3-trimethylammonium-propane (DSTAP), 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), and glucopyranosyl lipid adjuvant (GLA, also known as PHAD®) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL). Polysorbate 80 was purchased from J.T. Baker (San Francisco, CA). Poloxamer 188 was purchased from Spectrum Chemical (Gardena, CA). Saline solution (0.9% w/v) was purchased from Teknova (Hollister, CA). CpG ODN was obtained from Avecia (Milfrod, MA). Alhydrogel® ‘85’ and AdjuPhos® were purchased from E.M. Sergeant Pulp & Chemical Co. (Clifton, NJ). 3M-052 (S-36862) was synthesized by 3M Drug Delivery Systems. The chemical structure has been previously disclosed [16] (link).
1 2 dipalmitoyl 3 trimethylammonium propane dptap
Manufactured by Avanti Polar Lipids
Sourced in United States
1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP) is a cationic lipid commonly used in liposomal formulations and as a transfection reagent. It has a positively charged head group and two saturated fatty acid chains, which allows it to form stable lipid bilayers.
Automatically generated - may contain errors
Lab products found in correlation
Glucopyranosyl lipid adjuvant, by Avanti Polar Lipids (2 mentions)
1 2 dipalmitoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (2 mentions)
1 2 dipalmitoyl sn glycero 3 phosphocholine dppc, by Avanti Polar Lipids (2 mentions)
Saline solution, by Teknova (1 mentions)
Polysorbate 80, by Avantor (1 mentions)
Poloxamer 188, by Spectrum Chemical (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Imiquimod, by InvivoGen (1 mentions)
Lactic acid, by Merck Group (1 mentions)
1 2 dioleoyl sn glycero 3 phosphocholine dopc, by Avanti Polar Lipids (1 mentions)
Dspe peg2000, by Avanti Polar Lipids (1 mentions)
Dipalmitoylphosphatidylcholine dppc, by Avanti Polar Lipids (1 mentions)
A2780, by American Type Culture Collection (1 mentions)
Cfi plan apo lambda 100x oil, by Nikon (1 mentions)
Alexa 647 phalloidin, by Thermo Fisher Scientific (1 mentions)
Eclipse e600, by Nikon (1 mentions)
6 protocols using 1 2 dipalmitoyl 3 trimethylammonium propane dptap
1
Synthetic Lipids and Adjuvants for Vaccine Formulations
2
Lipid-based Adjuvant Formulations
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Milled imiquimod (IMQ) was purchased from Chemagis (Bnei Brak, Israel); imiquimod was also obtained from Invivogen (San Diego, CA). Glucopyranosyl lipid adjuvant (GLA), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2,dipalmitoyl-3-trimethylammonium-propane (DPTAP) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL). DOPC was also purchased from Lipoid (Newark, NJ). Cholesterol, and ammonium phosphate mono- and dibasic were purchased from J.T. Baker (San Francisco, CA). Lactic acid was purchased from Sigma-Aldrich (St. Louis, MO). Phosphate buffered saline (PBS) at pH 7.2 was purchased from Life Technologies (Grand Island, NY).
3
Preparation and Plasmid Loading of Lipid Microbubbles
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The MBs were fabricated by dissolving the dipalmitoyl-phosphatidyl-choline (DPPC) (Avanti Polar Lipids, AL, USA), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly(ethyleneglycol))-2000] (DSPE-PEG2000) (Avanti Polar Lipids) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP) (Avanti Polar Lipids) in chloroform at a molar ratio of 9:2:1, and then draining to generate a lipid film 30 (link). The film was then dissolved with 1 wt% glycerol-containing phosphate-buffered saline (PBS). Subsequently, the solution was degassed and refilled with perfluoropropane (C3F8). The MBs were formed by intense mechanical shaking using an agitator for 45 s. Finally, to separate from unreacted lipids, centrifugation was applied to the MBs at 6,000 rpm (2000 g) for 3 min.
Plasmid DNA (0-10000 ng) was gently mixed with 108 MBs for 30 min, and was centrifuged at 6,000 rpm (2000 g) for 1 min for separating unattached plasmid DNA. The normal MBs without plasmid DNA payload were prepared for comparison.
Plasmid DNA (0-10000 ng) was gently mixed with 108 MBs for 30 min, and was centrifuged at 6,000 rpm (2000 g) for 1 min for separating unattached plasmid DNA. The normal MBs without plasmid DNA payload were prepared for comparison.
4
Preparation and Characterization of Lipid-Based Nanomaterials
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The phospholipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, MW, 734.039 g/mol) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP) were purchased from Avanti Polar Lipids in USA. The phospholipids 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly(ethyleneglycol))-2000] (DSPE-PEG2000-Biotin, MW, 3,016.781 g/mol) and Biotin-PEG(2k)-SS-NHS were purchased from Xi’an Ruixi Biological Technology Co. in China. Perfluoropropane (C3F8) gas was purchased from the R&D Center for Specialty Gases at the Research Institute of Physical and Chemical Engineering of Nuclear Industry (Beijing, China). FITC-Avidin was purchased from Solarbio Inc (Beijing, China). The human ovarian cancer cells A2780 were from the American Type Culture Collection.
5
Preparation of Mica-Supported Lipid Bilayer
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We tested several surfaces, including glass and mica, to be used during the HS-AFM experiments. We found that mica-SLB is the best one for actin and TF imaging as well for TF–myosin II interaction. Mica disks (1.5 mm in diameter; RIBM) were glued on the top of glass cylindrical sample stage and freshly cleaved before depositing lipids. The positively charged lipid bilayer was prepared according to Uchihashi et al. (54 (link)) with several modifications. Briefly, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) (Avanti Polar Lipids) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP) (Avanti Polar Lipids) in 100% chloroform were mixed at a weight ratio 9:1 or 9.5:0.5, dried from organic solvent with warming up to 45 °C, and dissolved to 2 mg/mL by solution, containing 50 mM KCl, 4 mM MgCl2, 0.5 mM EGTA, 25 mM imidazole-HCl, 1 mM DTT, pH 6.0 (buffer A). The mix then was vortexed, sonicated to disperse the lipids for 2 to 4 min, and subjected to snap freezing in liquid N2 and thawing (5 cycles). Lipids were then aliquoted and stored at −80 °C. Before use, the aliquot was diluted five times by 10 mM MgCl2 (to 0.4 mg/mL lipid solution) and sonicated again for 2 min. Lipids were placed on the top of freshly cleaved mica and incubated at room temperature in the humid chamber to prevent drying the lipids for at least 30 min or overnight.
6
Visualization of Actin-HMM-GFP Binding
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Binding of HMM-GFP to actin filaments was observed as follows. G-actin was polymerized in NF-buffer (50 mM KCl, 2 mM MgCl 2 , 0.5 mM EGTA, 1 mM DTT, 20 mM PIPES, pH 6.5) containing 0.2 mM ATP for 2 h at 22°C. Actin filaments and Alexa 647 phalloidin (Invitrogen)
were mixed at a molar ratio of 20:1 and incubated overnight on ice. The surface of coverslips was covered with a positively charged lipid bilayer and was used to construct flow chambers as described previously (18) , except that the weight ratio of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC;
Avanti Polar Lipids, Alabaster, AL) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP;
Avanti Polar Lipids) was 17:3 (14) . Alexa 647 phalloidin-stabilized actin filaments diluted in NF-ATP buffer (NF buffer containing 0.5 µM ATP) were introduced into the flow chamber to loosely bind on the positively charged lipid monolayer. HMM-GFP and Rng2CHD diluted in NF-ATP buffer were then introduced to the flow chamber. Alternatively, NF-ATP buffer in the above procedures was replaced with NF buffer for the assays in the nucleotide-free state. Fluorescence of Alexa 647 and GFP was imaged with a fluorescence microscope (ECLIPSE E600, Nikon, Tokyo, Japan) equipped with an ARUGUS-HiSCA system (Hamamatsu Photonics, Hamamatsu, Japan). Images were captured using a 100x objective lens (CFI Plan Apo Lambda 100x Oil, NA 1.45; Nikon).
were mixed at a molar ratio of 20:1 and incubated overnight on ice. The surface of coverslips was covered with a positively charged lipid bilayer and was used to construct flow chambers as described previously (18) , except that the weight ratio of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC;
Avanti Polar Lipids, Alabaster, AL) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP;
Avanti Polar Lipids) was 17:3 (14) . Alexa 647 phalloidin-stabilized actin filaments diluted in NF-ATP buffer (NF buffer containing 0.5 µM ATP) were introduced into the flow chamber to loosely bind on the positively charged lipid monolayer. HMM-GFP and Rng2CHD diluted in NF-ATP buffer were then introduced to the flow chamber. Alternatively, NF-ATP buffer in the above procedures was replaced with NF buffer for the assays in the nucleotide-free state. Fluorescence of Alexa 647 and GFP was imaged with a fluorescence microscope (ECLIPSE E600, Nikon, Tokyo, Japan) equipped with an ARUGUS-HiSCA system (Hamamatsu Photonics, Hamamatsu, Japan). Images were captured using a 100x objective lens (CFI Plan Apo Lambda 100x Oil, NA 1.45; Nikon).
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