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96
Bio-Rad precision plus protein unstained standards
Fig. 5. Role of fluidity for Hla–SM interaction. Liposomes containing 40% cholesterol and variable fractions of two types of sphingomyelin (17 μM in the outer leaflet) were incubated for 40 s or 45 min at 20 °C or 37 °C with 4 μM pyrene labelled Hla. The rest of the lipid consists of ePE and bPS in a ratio of 1: 1. Panel A: Relative excimer fluorescence as obtained for pseudo-ternary and pseudo-binary mixtures (10, 20 and 40% SM with 40% cholesterol and 1: 1 ePE/bPS) and binary mixtures (60% SM with 40% cholesterol, indicated by a star). Samples were diluted to 0.1 μM directly before measurement. Bars indicated liposomes containing eSM, symbols (squares, triangles, circles, diamond) represent liposomes containing OSM as Hla-binding lipid. For comparison, fluorescence levels measured in absence of liposomes (“toxin”) are depicted. After incubation, aliquots of the samples were subjected to 10% SDS–PAGE without heating. A gel showing part of the samples (45 min, 37 °C) is depicted in panel B; the numbers represent the fraction of oligomers based on a densitometric analysis (Quantity One, Bio-Rad, München, Germany). The results for all samples are depicted in panel C, with bars indicating liposomes containing eSM and symbols indicating liposomes containing OSM. Incubation time and temperature was as given in the legend. Marker: <t>Precision</t> <t>Plus</t> <t>Protein</t> <t>unstained</t> <t>Standards</t> (Bio-Rad, München, Germany).
Precision Plus Protein Unstained Standards, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Bio-Rad precision plus protein standard
Fig. 5. Role of fluidity for Hla–SM interaction. Liposomes containing 40% cholesterol and variable fractions of two types of sphingomyelin (17 μM in the outer leaflet) were incubated for 40 s or 45 min at 20 °C or 37 °C with 4 μM pyrene labelled Hla. The rest of the lipid consists of ePE and bPS in a ratio of 1: 1. Panel A: Relative excimer fluorescence as obtained for pseudo-ternary and pseudo-binary mixtures (10, 20 and 40% SM with 40% cholesterol and 1: 1 ePE/bPS) and binary mixtures (60% SM with 40% cholesterol, indicated by a star). Samples were diluted to 0.1 μM directly before measurement. Bars indicated liposomes containing eSM, symbols (squares, triangles, circles, diamond) represent liposomes containing OSM as Hla-binding lipid. For comparison, fluorescence levels measured in absence of liposomes (“toxin”) are depicted. After incubation, aliquots of the samples were subjected to 10% SDS–PAGE without heating. A gel showing part of the samples (45 min, 37 °C) is depicted in panel B; the numbers represent the fraction of oligomers based on a densitometric analysis (Quantity One, Bio-Rad, München, Germany). The results for all samples are depicted in panel C, with bars indicating liposomes containing eSM and symbols indicating liposomes containing OSM. Incubation time and temperature was as given in the legend. Marker: <t>Precision</t> <t>Plus</t> <t>Protein</t> <t>unstained</t> <t>Standards</t> (Bio-Rad, München, Germany).
Precision Plus Protein Standard, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
Rad Laboratories Inc precision plus protein standard (unstained)
Fig. 5. Role of fluidity for Hla–SM interaction. Liposomes containing 40% cholesterol and variable fractions of two types of sphingomyelin (17 μM in the outer leaflet) were incubated for 40 s or 45 min at 20 °C or 37 °C with 4 μM pyrene labelled Hla. The rest of the lipid consists of ePE and bPS in a ratio of 1: 1. Panel A: Relative excimer fluorescence as obtained for pseudo-ternary and pseudo-binary mixtures (10, 20 and 40% SM with 40% cholesterol and 1: 1 ePE/bPS) and binary mixtures (60% SM with 40% cholesterol, indicated by a star). Samples were diluted to 0.1 μM directly before measurement. Bars indicated liposomes containing eSM, symbols (squares, triangles, circles, diamond) represent liposomes containing OSM as Hla-binding lipid. For comparison, fluorescence levels measured in absence of liposomes (“toxin”) are depicted. After incubation, aliquots of the samples were subjected to 10% SDS–PAGE without heating. A gel showing part of the samples (45 min, 37 °C) is depicted in panel B; the numbers represent the fraction of oligomers based on a densitometric analysis (Quantity One, Bio-Rad, München, Germany). The results for all samples are depicted in panel C, with bars indicating liposomes containing eSM and symbols indicating liposomes containing OSM. Incubation time and temperature was as given in the legend. Marker: <t>Precision</t> <t>Plus</t> <t>Protein</t> <t>unstained</t> <t>Standards</t> (Bio-Rad, München, Germany).
Precision Plus Protein Standard (Unstained), supplied by Rad Laboratories Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Fig. 5. Role of fluidity for Hla–SM interaction. Liposomes containing 40% cholesterol and variable fractions of two types of sphingomyelin (17 μM in the outer leaflet) were incubated for 40 s or 45 min at 20 °C or 37 °C with 4 μM pyrene labelled Hla. The rest of the lipid consists of ePE and bPS in a ratio of 1: 1. Panel A: Relative excimer fluorescence as obtained for pseudo-ternary and pseudo-binary mixtures (10, 20 and 40% SM with 40% cholesterol and 1: 1 ePE/bPS) and binary mixtures (60% SM with 40% cholesterol, indicated by a star). Samples were diluted to 0.1 μM directly before measurement. Bars indicated liposomes containing eSM, symbols (squares, triangles, circles, diamond) represent liposomes containing OSM as Hla-binding lipid. For comparison, fluorescence levels measured in absence of liposomes (“toxin”) are depicted. After incubation, aliquots of the samples were subjected to 10% SDS–PAGE without heating. A gel showing part of the samples (45 min, 37 °C) is depicted in panel B; the numbers represent the fraction of oligomers based on a densitometric analysis (Quantity One, Bio-Rad, München, Germany). The results for all samples are depicted in panel C, with bars indicating liposomes containing eSM and symbols indicating liposomes containing OSM. Incubation time and temperature was as given in the legend. Marker: Precision Plus Protein unstained Standards (Bio-Rad, München, Germany).

Journal: Biochimica et biophysica acta

Article Title: Lipid and phase specificity of α-toxin from S. aureus.

doi: 10.1016/j.bbamem.2013.04.005

Figure Lengend Snippet: Fig. 5. Role of fluidity for Hla–SM interaction. Liposomes containing 40% cholesterol and variable fractions of two types of sphingomyelin (17 μM in the outer leaflet) were incubated for 40 s or 45 min at 20 °C or 37 °C with 4 μM pyrene labelled Hla. The rest of the lipid consists of ePE and bPS in a ratio of 1: 1. Panel A: Relative excimer fluorescence as obtained for pseudo-ternary and pseudo-binary mixtures (10, 20 and 40% SM with 40% cholesterol and 1: 1 ePE/bPS) and binary mixtures (60% SM with 40% cholesterol, indicated by a star). Samples were diluted to 0.1 μM directly before measurement. Bars indicated liposomes containing eSM, symbols (squares, triangles, circles, diamond) represent liposomes containing OSM as Hla-binding lipid. For comparison, fluorescence levels measured in absence of liposomes (“toxin”) are depicted. After incubation, aliquots of the samples were subjected to 10% SDS–PAGE without heating. A gel showing part of the samples (45 min, 37 °C) is depicted in panel B; the numbers represent the fraction of oligomers based on a densitometric analysis (Quantity One, Bio-Rad, München, Germany). The results for all samples are depicted in panel C, with bars indicating liposomes containing eSM and symbols indicating liposomes containing OSM. Incubation time and temperature was as given in the legend. Marker: Precision Plus Protein unstained Standards (Bio-Rad, München, Germany).

Article Snippet: Marker: Precision Plus Protein unstained Standards (Bio-Rad, 0.037 s−1 and 0.108 s−1) but the overall oligomerisation level is similar for 4 °C, 22 °C and 37 °C.

Techniques: Liposomes, Incubation, Binding Assay, Comparison, SDS Page, Marker