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1 2 dioleoyl sn glycero 3 phosphoethanolamine n lissamine rhodamine b sulfonyl rhod pe

Manufactured by Avanti Polar Lipids
Sourced in United States

1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rhod-PE) is a fluorescent lipid probe. It consists of a phosphatidylethanolamine headgroup with a lissamine rhodamine B sulfonyl fluorophore attached to the ethanolamine nitrogen. Rhod-PE can be used to label and visualize lipid membranes and vesicles.

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5 protocols using 1 2 dioleoyl sn glycero 3 phosphoethanolamine n lissamine rhodamine b sulfonyl rhod pe

1

Fluorescent Lipid Labeling Protocol

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Peptide synthesis reagents and Fmoc-protected amino acids were purchased from Iris Biotech (Marktredwitz, Germany) or Merck Biosciences (Darmstadt, Germany). Chemicals for peptide synthesis were obtained from Merck (Darmstadt, Germany) or Biosolve (Valkenswaard, Netherlands), and solvents for HPLC purification were purchased from Fischer Scientific (Geel, Belgium). 4-Chloro-7-nitrobenzofurazan (NBD-Cl) was purchased from VWR (Bruchsal, Germany). The fluorescent probes 8-amino-naphtalene-1,3,6-trisulfonic acid sodium salt (ANTS) and p-xylene-bis-pyridinium bromide (DPX) were obtained from Invitrogen-Molecular Probes (Karlsruhe, Germany). Fluorescently labelled 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rhod-PE) was obtained from Avanti Polar Lipids (Alabaster, AL, USA). The lipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG), 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC), and 1,2-dimyristoyl-sn-glycero-3-phosphatidylglycerol (DMPG) were purchased from NOF (Grobbendonk, Belgium).
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2

Synthesis and Labeling of Lipids

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15N-labelled amino acids were purchased from Cambridge Isotope Laboratories (Andover, MA, USA) and were Fmoc-protected using Fmoc-Cl as described previously [33 (link)]. The lipids 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphatidylglycerol (DMPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG), were purchased from NOF Europe (Grobbendonk, Belgium). The fluorescent lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rhod-PE) were obtained from Avanti Polar Lipids. The fluorescent probes 8-amino-naphtalene-1,3,6-trisulfonic acid sodium salt (ANTS) and p-xylene-bis(pyridinium)bromide (DPX) were obtained from Invitrogen-Molecular Probes (Karlsruhe, Germany).
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3

PEG-PBD and PEG-PPO Copolymer Synthesis

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Poly(ethylene glycol) (900)-polybutadiene (1800) copolymer (denoted PEG-PBD, MW 2700 Da) and Poly(ethylene glycol) (450)-poly(propylene oxide) (1400) (denoted PEG-PPO, MW 1850 Da) were purchased from Polymer Source (Dorval, Quebec, Canada). 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(Lissamine rhodamine B sulfonyl) (Rhod-PE) was purchased from Avanti Polar Lipids, Inc. Cu/Zn Superoxide dismutase (SOD; MW 32500) from bovine erythrocytes was from Calbiochem (EMD Millipore, Billerica, MA). Fluorescein isothiocyanate (FITC) was purchased from Thermo Fisher Scientific. IRDye 800CW NHS Ester was purchased from LI-COR, Inc. All other chemicals were used as received. All of the buffer solutions were prepared with deionized water.
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4

Peptide Synthesis and Lipid Preparation

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Peptide synthesis reagents and Fmoc-protected amino acids were purchased from Merck Biosciences (Darmstadt, Germany) and/or Iris Biotech (Marktretwitz, Germany). 15N-labelled amino acids were purchased from Cambridge Isotope Laboratories (Andover, MA, USA) and were Fmoc-protected using Fmoc-Cl as described previously69 . Solvents for peptide synthesis were purchased from Merck (Darmstadt, Germany) or from Biosolve (Valkenswaard, Netherlands) and solvents for HPLC purification were obtained from Fischer Scientific (Geel, Belgium). The lipids 1,2-dierucoyl-sn-glycero-3-phosphatidylcholine (DErPC), 1,2-dierucoyl-sn-glycero-3-phosphatidylglycerol (DErPG), 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphatidylglycerol (DMPC), 1,2-dimyristoleoyl-sn-glycero-3-phosphatidylcholine (DMoPC), 1,2-dioleoyl-sn-glycero-3-phosphatidylglycerol (DOPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rhod-PE) were obtained from Avanti Polar Lipids (Alabaster, USA). The fluorescent probes 8-amino-naphtalene-1,3,6-trisulfonic acid sodium salt (ANTS) and pxylenebis(pyridinium)bromide (DPX) were obtained from Invitrogen - Molecular Probes (Karlsruhe, Germany).
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5

PLA2-Mediated Liposomal Delivery

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Thioetheramide-PC was purchased from Cayman Chemical (Ann Arbor, MI). HSPC, DSPE-PEG2000, DOTAP, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(Lissamine rhodamine B sulfonyl) (Rhod-PE), DSPE-PEG2000-Cy7, and NBD-PC were purchased from Avanti Polar Lipids Inc. sPLA2 enzyme from Naja mossambica was purchased from Sigma-Aldrich Co. Rabbit polyclonal antibody to sPLA2-IIa, p-P65, and p-P100 antibodies were purchased from Abcam. Bovine synovial fluid was obtained from Lampire Biological Laboratories Inc. All other chemicals were used as received. All of the buffer solutions were prepared with deionized water.
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