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Topfluor lysopc

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TopFluor LysoPC is a lipid probe that incorporates a fluorescent TopFluor label onto the lyso-phosphatidylcholine (lysoPC) headgroup. It can be used to visualize and track the localization of lysoPC in biological systems.

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Lab products found in correlation

Texas red 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine triethylammonium salt tr dhpe, by Thermo Fisher Scientific (2 mentions) Direct q 3uv r, by Merck Group (2 mentions) Sodium chloride (nacl), by Merck Group (2 mentions) Bodipy fl, by Avanti Polar Lipids (2 mentions) Lsm 700 confocal microscope, by Zeiss (2 mentions) Phosphate buffer, by Merck Group (1 mentions) Survanta, by Abbvie (1 mentions) Lysopc, by Avanti Polar Lipids (1 mentions) Methylthiazolyldiphenyl tetrazolium bromide mtt, by Merck Group (1 mentions) Protease inhibitor cocktail p8340, by Merck Group (1 mentions) Dspe peg2000, by Avanti Polar Lipids (1 mentions) Topfluor pc, by Avanti Polar Lipids (1 mentions) N methoxy polyethylene glycol 2000, by Avanti Polar Lipids (1 mentions) Bodipy tr c5 ceramide, by Thermo Fisher Scientific (1 mentions) Prism 6, by GraphPad (1 mentions) Tb fluorescent stain kit m, by BD (1 mentions) Alexa 647, by Thermo Fisher Scientific (1 mentions) Bodipy 493 503, by Thermo Fisher Scientific (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) β lactoglobulin, by Merck Group (1 mentions)

6 protocols using topfluor lysopc

1

Mitochondrial Lipid Dynamics in COS-1 Cells

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COS-1 cells were seeded in four-well chambered cover glass (Cellvis, cat# C4-1.5H-N) at a density of 1.2 × 105 cells per well, and mitochondria were labeled with MitoTracker™ Red CMXRos 18 h later when cells are adherent. Chambered cover glass was placed on ice 6 h after mitochondria staining, washed with KHM buffer [110 mM potassium acetate, 20 mM HEPES (pH 7.4), 2 mM MgCl2] twice and permeabilized with 20 µM digitonin (in KHM buffer) on ice for 4 min. Permeabilized cells were washed with KHM buffer twice and incubated with warm DMEM medium supplemented with either 0.5% (v/v) methanol or 5 µM TopFluor LysoPC (Avanti Polar Lipids, cat#810284, dissolved in methanol, 0.5% (v/v)) for 10 min at 37°C. Cells were then washed twice with warm DMEM and examined under an LSM 800 confocal microscope.
Li X., Anderson D.E., Chang Y.Y., Jarnik M, & Machner M.P. (2022). VpdC is a ubiquitin-activated phospholipase effector that regulates Legionella vacuole expansion during infection. Proceedings of the National Academy of Sciences of the United States of America, 119(48), e2209149119.
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2

Lipid Characterization and Fluorescent Labeling

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Lysopalmitoylphosphatidylcholine (LysoPC) was purchased from Avanti Polar Lipids (Huntsville, AL) and used as received. LysoPC has a critical micelle concentration (CMC) of ~ 6 μM and a minimum surface tension of ~ 36 mN/m 2 at the CMC and higher concentrations. Water with a resistivity of 18.2 MΩ·cm at 25 °C was purified with a Millipore Direct Q 3UV-R (Millipore, Billerica, MA) system. Sodium chloride (NaCl), and phosphate buffer were purchased in powder form from Sigma-Aldrich (Saint Louis, MO, USA), and used to prepare phosphate buffered saline (PBS) solutions (150 mM NaCl and pH 7.0). Survanta (AbbVie Inc., IL, USA) was diluted to 2 mg/mL in PBS before use. The lipid dye Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt, (TR-DHPE) was purchased from Life Technologies Corporation, CA, USA and used as received. 1-(dipyrrometheneboron difluoride)undecanoyl-2-hydroxy-sn-glycero-3-phosphocholine, or TopFluor Lyso PC, a green fluorescent derivative of LysoPC with spectral properties similar to Bodipy-FL was purchased from Avanti Polar Lipids and used as received.
Barman S., Davidson M.L., Walker L.M., Anna S.L, & Zasadzinski J.A. (2020). Inflammation Product Effects on Dilatational Mechanics Can Trigger the Laplace Instability and Acute Respiratory Distress Syndrome. Soft matter, 16(29), 6890-6901.
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3

Lipid Nanoparticle Isolation and Characterization

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PM was extracted from mouse PLTs according to our previous method 27 (link). DPPC, MSPC,1-palmitoyl-2-(dipyrrometheneborondifluoride)undecanoyl-sn-glycero-3-phosphoethanolamine (TopFluor®PC, 495 nm/503 nm), 1-(dipyrrometheneboron difluoride)undecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (TopFluor® Lyso PC) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000) were obtained from Avanti Polar Lipids Inc. 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine (DiR) iodide (748 nm/780 nm) was purchased from AAT Bioquest Inc. Doxorubicin, Protease inhibitor cocktail (P8340) and methylthiazolyldiphenyl-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich. Deionized water (18.2 MΩ cm) from a Milli-Q purification system was used in all preparations.
Wu D., Jin X., Wang X., Ma B., Lou C., Qu H., Zheng J., Zhang B., Yan X., Wang Y, & Jing L. (2019). Engineering temperature-sensitive plateletsomes as a tailored chemotherapy platform in combination with HIFU ablation for cancer treatment. Theranostics, 9(13), 3966-3979.
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4

Lipid Dye Staining of Malaria Parasites

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Bodipy-TR-C5-ceramide (Invitrogen) staining was performed using a concentration of 2.5 μM (stock 5 μM) in RPMI as previously described [46 (link)] in ring and trophozoites of condΔEXP1 parasites and condΔEXP1 expressing EXP2-GFP after 1 cycle ± rapalog. For Lyso PC labelling, TopFluor LysoPC (Avanti Polar Lipids, Alabaster, AL) (1 mM stock in methanol) was resuspended in PBS to a final concentration of 20 μM, added to ring and trophozoite stages of control and ΔEXP1 parasites, and incubated for 15 minutes at 37°C. All microscopy images of the lipid dye stained parasites were recorded with the same acquisition settings and exposure time. Number of protrusions in each parasite were counted, and data were analyzed with Graph Pad Prism 6.07 (Graph Pad Software, http://www.graphpad.com).
Mesén-Ramírez P., Bergmann B., Tran T.T., Garten M., Stäcker J., Naranjo-Prado I., Höhn K., Zimmerberg J, & Spielmann T. (2019). EXP1 is critical for nutrient uptake across the parasitophorous vacuole membrane of malaria parasites. PLoS Biology, 17(9), e3000473.
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5

Fluorescent Lipid Imaging in Dictyostelium

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Bodipy 493/503, Bodipy 558/568 (BodipyC12) and Vbyrant Ruby were purchased from Thermo Scientific. LD540 was a gift from Prof. Christoph Thiele (Bonn, Germany). The AuramineO staining was performed using the TB Fluorescent Stain Kit M (Becton Dickinson). The p80-antibody [59 (link)] was obtained from the lab of Dr. Pierre Cosson (University of Geneva). Goat-anti mouse IgG coupled to Alexa488 or Alexa647 (Thermo Scientific) were used as secondary antibodies. Topfluor-LysoPC, Topfluor-FAs (C11) and Topfluor-TAGs (18:1, 18:1, C11) were purchased from Avanti Polar Lipids.
Dictyostelium was fixed with 4% PFA/picric acid or ultra cold methanol as described previously [60 (link)]. Staining with Bodipy 493/503 at a final concentration of 20 μM was performed in parallel with the secondary antibody. Images were recorded with a Zeiss LSM700 confocal microscope using a 63× 1.4 NA or 100× 1.4 NA oil immersion objective.
Barisch C, & Soldati T. (2017). Mycobacterium marinum Degrades Both Triacylglycerols and Phospholipids from Its Dictyostelium Host to Synthesise Its Own Triacylglycerols and Generate Lipid Inclusions. PLoS Pathogens, 13(1), e1006095.
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6

Lipid-Protein Interactions in Biomembranes

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Lysophosphatidylcholine (LysoPC, MW=495.63 g/mol, CMC = 6 μM), a saturated 16 carbon single-tail lipid, was purchased in powder form from Anatrace and was used as received. Dipalmitoylphosphatidylcholine (DPPC, MW=734.04 g/mol), a saturated 16 carbon double-tailed phospholipid, was purchased in powder form from Avanti Lipids and was used as received. β-lactoglobulin (18.4 kDa) was purchased in powder form from Sigma Aldrich and was used as received. Water with a resistivity of 18.2 MΩ·cm at 25 °C was purified with a Millipore Direct Q 3UV-R (Millipore, Billerica, MA) system. Sodium chloride (NaCl) and phosphate buffer were purchased in powder form from Sigma-Aldrich (Saint Louis, MO, USA) and used to prepare phosphate buffered saline (PBS) solutions (150 mM NaCl and pH 7.0). The lipid dye, Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt, (TR-DHPE) was purchased from Life Technologies Corporation, CA, USA and used as received. 1-(dipyrrometheneboron-difluoride)undecanoyl-2-hydroxy-sn-glycero-3-phosphocholine, or TopFluor Lyso PC, a green fluorescent derivative of LysoPC with spectral properties similar to Bodipy-FL was purchased from Avanti Polar Lipids.
Reinbothe S., Reinbothe C., Neumann D, & Apel K. (1996). A plastid enzyme arrested in the step of precursor translocation in vivo.. Proceedings of the National Academy of Sciences of the United States of America, 93(21), 12026-12034.
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