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the fret software package  (Evident Corporation)

 
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    Evident Corporation the fret software package
    The Fret Software Package, supplied by Evident Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/fret+software+package/the+fret+software+package/pmc02785618-72-7-11
    Average 90 stars, based on 1 article reviews
    the fret software package - by Bioz Stars, 2026-09
    90/100 stars

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    Article Title: The Tec Family Kinase Itk Exists as a Folded Monomer in Vivo
    Article Snippet: For FRET analysis, cells were imaged 36 h following transfection into 293T cells, using an Olympus Fluoview confocal microscope. .. The normalized FRET was calculated using the FRET software package from Olympus. ..



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    Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, <t>FRET</t> <t>analysis</t> of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.
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    Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, <t>FRET</t> <t>analysis</t> of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.
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    Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, <t>FRET</t> <t>analysis</t> of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.
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    Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, <t>FRET</t> <t>analysis</t> of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.
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    Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, FRET analysis of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.

    Journal: The Journal of Neuroscience

    Article Title: MCU Interacts with Miro1 to Modulate Mitochondrial Functions in Neurons

    doi: 10.1523/JNEUROSCI.0504-18.2018

    Figure Lengend Snippet: Domain mapping of the interaction between MCU and Miro1. A, Mutations on the dime motif of MCU (257EYSW260 to 257AAAA260 or 261DIME264 to 261AAAA264) did not affect the interaction between MCU and Miro1. B, MCU did not interact with Myc-Miro1 without transmembrane domain, but still immunoprecipitated Myc-Miro1 lacking 3 aa (616KQR618). C, FRET analysis of Myc-Miro1 and Flag-MCU showed a significantly higher FRET efficiency compared with that of TOMM20-Myc and Flag-MCU. Mutation on the dime motif of MCU (257EYSW260 to 257AAAA260) did not affect Myc-Miro1 binding, while elimination of MCU's N-terminal domain [Flag-MCU(Δ2–57)] or adding Ca2+ significantly reduced interaction with Myc-Miro1. N = Flag-MCU + TOMM20-Myc: 72 mitochondria. Flag-MCU + Myc-Miro1: 78 mitochondria, Flag-MCU (257EYSW260 to 257AAAA260) + Myc-Miro1: 25 mitochondria, Flag-MCU(Δ2–57) + Myc-Miro1: 23 mitochondria. Scale bar, 1 μm. *p < 0.0001. Tukey's box plot profile is shown. D, Ca2+ elevation alters MCU–Miro1 interaction. MCU expression vector dually tagged in the N-terminus (Flag) and the C-terminus (sfGFP), respectively, was prepared and used to investigate localization of these tags after treating cells with 2 μm ionomycin for 10 min. The R2 correlation between Flag and sfGFP was compared between paired samples with or without addition of 2 μm ionomycin. N = 5 experiments, *p = 0.0171. E, HEK cells transfected with MCU-Flag were treated with 2 μm ionomycin as shown above and then blotted for Flag. Graph shows the ratio of the full-length MCU band (top band) to the processed MCU (bottom band). N = 7 experiments, **p = 0.0094. Tukey's boxplot profile is shown.

    Article Snippet: All FRET efficiencies were calculated and normalized using the Zeiss FRET analysis software package.

    Techniques: Immunoprecipitation, Mutagenesis, Binding Assay, Expressing, Plasmid Preparation, Transfection