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Bodipy tr c5 ceramide

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BODIPY TR C5-ceramide is a fluorescent lipid analog that can be used as a marker for cellular lipids and membranes. It contains a BODIPY fluorescent dye conjugated to a C5-ceramide moiety.

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Lab products found in correlation

Ix83 inverted fluorescence microscope, by Olympus (1 mentions) β casein from bovine milk, by Merck Group (1 mentions) 1 2 distearoyl sn glycero 3 phosphoethanolamine n biotinyl polyethylene glycol 2000, by Avanti Polar Lipids (1 mentions) Prism 6, by GraphPad (1 mentions) Topfluor lysopc, by Avanti Polar Lipids (1 mentions) Chlorpromazine, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Sucrose, by Merck Group (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Er tracker red, by Thermo Fisher Scientific (1 mentions) Filipin, by Merck Group (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) 1 2 distearoyl sn glycero 3 phosphoethanolamine n biotinyl polyethylene glycol 2 000 dspe peg 2 000 biotin, by Avanti Polar Lipids (1 mentions) β casein, by Merck Group (1 mentions) L α phosphatidylinositol 4 5 bisphosphate pi 4 5 p2, by Avanti Polar Lipids (1 mentions)

Spelling variants of this product

BODIPY (264 citations) BODIPY TR Ceramide (43 citations) BODIPY-ceramide (9 citations) BODIPY C5 Ceramide (2 citations)

6 protocols using bodipy tr c5 ceramide

1

Live-cell imaging of T. gondii infection

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HeLa cells were seeded on 384-well glass bottom plates, transfected with Arf1-GFP, and infected with an untagged T. gondii RH-88 strain as described above. At 24 hpi, cells were incubated with 5 µM BODIPY TR C5-ceramide complexed to BSA (Invitrogen; Cat# B34400) in FBS-free DMEM. Nuclei were stained with Hoechst (1:2,000 in PBS) for 5 minutes before live cell microscopy at 48 hpi.
Schroeder E.A., Toro-Moreno M., Raphemot R., Sylvester K., Colón I.C, & Derbyshire E.R. (2024). Toxoplasma and Plasmodium associate with host Arfs during infection. mSphere, 9(3), e00770-23.
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2

Localization of TcPho91-GFP in Trypanosoma Epimastigotes

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Epimastigotes overexpressing TcPho91-GFP and maintained either in LIT or SDM-79 media were collected by centrifugation in the exponential phase of growth and resuspended at a density of 1×107 parasites/ml in LIT or SDM79 media containing 5 μM of BODIPY-TR C5 ceramide (Invitrogen). After 1 h of incubation at 28°C, the cells were collected by centrifugation at 1,600 × g, and washed twice in PBS, pH 7.4. Labeled epimastigotes were then placed in bottom-glass petri dishes previously treated with poly-lysine. Cells were allowed to settle in the dish for 2 min and loose parasites were washed away with isosmotic buffer. Live cell imaging was done in isosmotic conditions in an Olympus IX-83 inverted fluorescence microscope and processed with CellSense Dimensions software.
Jimenez V, & Docampo R. (2015). TcPho91 is a contractile vacuole phosphate sodium symporter that regulates phosphate and polyphosphate metabolism in Trypanosoma cruzi. Molecular microbiology, 97(5), 911-925.
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3

Lipid Extraction and Labeling for Fluorescence Imaging

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Total brain lipid extract (131101P), brain L-α-PI(4,5)P2 (840046P), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethyleneglycol)-2000] (880129P) were purchased from Avanti Polar Lipids (Alabaster, AL). BODIPY-TR-C5-ceramide, (BODIPY-TR ceramide, D7540), BODIPYFL C5-hexadecanoyl phosphatidylcholine (D3803), and Alexa Fluor 488 (AX488) C5-Maleimide were purchased from Invitrogen (Carlsbad, CA). GloPIPs BODIPY TMR-PtdIns(4,5)P2, C16 (C45M16a) was purchased from Echelon Biosciences (Salt Lake City, UT). β-Casein from bovine milk (>98% pure, C6905) and other reagents were purchased from Sigma-Aldrich (St. Louis, MO).
Jarin Z., Tsai F.C., Davtyan A., Pak A.J., Bassereau P, & Voth G.A. (2019). Unusual Organization of I-BAR Proteins on Tubular and Vesicular Membranes. Biophysical Journal, 117(3), 553-562.
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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

Cellular Processes Inhibition Assay

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Bovine serum albumin and all cellular process inhibitors (protease inhibitor E64, chlorpromazine (CPZ, a cationic amphiphilic inhibitor acts on clathrin-coated pits), sucrose (hypertonic media that interferes with clathrin and adaptor protein interactions), bafilomycin A1 and filipin) were purchased from Sigma USA). ER-Tracker™Red, BODIPY TR C5-ceramide, LysoTracker, mouse anti-Rad5, Hoechst, propidium iodide, Alexa Fluor 488 and carboxylic acid succinimidyl ester were obtained from Molecular Probes (USA).
Chaiyadet S., Smout M., Johnson M., Whitchurch C., Turnbull L., Kaewkes S., Sotillo J., Loukas A, & Sripa B. (2015). Excretory/secretory products of the carcinogenic liver fluke are endocytosed by human cholangiocytes and drive cell proliferation and IL6 production. International journal for parasitology, 45(12), 773-781.
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6

Lipid Extraction and Labeling Protocol

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Brain total lipid extract, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethyleneglycol)-2000] (DSPE-PEG(2000)-biotin), and L-α- phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) were purchased from Avanti Polar Lipids. BODIPY-TR-C5-ceramide was obtained from Molecular Probes. All reagents used to make buffers and β-casein from bovine milk (>99%) were purchased from Sigma-Aldrich.
Renard H.F., Simunovic M., Lemière J., Boucrot E., Garcia-Castillo M.D., Arumugam S., Chambon V., Lamaze C., Wunder C., Kenworthy A.K., Schmidt A.A., McMahon H.T., Sykes C., Bassereau P, & Johannes L. (2014). ENDOPHILIN-A2 FUNCTIONS IN MEMBRANE SCISSION IN CLATHRIN-INDEPENDENT ENDOCYTOSIS. Nature, 517(7535), 493-496.
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