Recombinant:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Luciferase:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Membrane:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Protein Purification:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Cryo-EM Sample Prep:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Software:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
Microscopy:Article Title: Structure of human Frizzled5 by fiducial-assisted cryo-EM supports a heterodimeric mechanism of canonical Wnt signaling
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.58464 11 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein DRPB_Fz8 Dang et al., 2019 (PMID:31086346) For signaling assay Commercial assay or kit Luciferase assay kit Promega E152A For signaling assay Chemical compound, drug n-Dodecylb-D-Maltoside (DDM) Anatrace D310S Membrane protein purification Chemical compound, drug Lauryl Maltose Neopentyl Glycol (LMNG) Anatrace NG310 Membrane protein purification Chemical compound, drug Glyco-Diosgenin (GDN) Anatrace GDN101 Membrane protein purification Chemical compound, drug Cholesterol Hemisuccinate tris Salt (CHS) Anatrace CH210 Membrane protein purification Chemical compound, drug Digitonin EMD Millipore 300410 Cryo-EM specimen freezing additive Software, algorithm DigitalMicrograph Gatan Microscope alignment and cryo-EM data collection Software, algorithm serialEM https://bio3d. colorado.edu/ SerialEM/ (PMID:16182563) version 3.6 Cryo-EM data collection Software, algorithm MotionCor2 https://emcore. ucsf.edu/ucsfmotioncor2/ (PMID:28250466) Motion correction of cryo-EM movies Software, algorithm cryoSPARC https://cryosparc.com/ (PMID:28165473) version 2.12.14 Cryo-EM data processing Software, algorithm Coot https://www2. mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ (PMID:20383002) Structure model building Software, algorithm Isolde https://isolde. cimr.cam.ac.uk/ (PMID:29872003) Structure model building/ refinement Software, algorithm Phenix suite https://www. phenix-online.org/ (PMID:20124702) Structure refinement Software, algorithm UCSF Chimera https://www.cgl. ucsf.edu/chimera/ (PMID:15264254) Initial homology model docking Software, algorithm UCSF ChimeraX https://www. rbvi.ucsf.edu/ chimerax/ (PMID:28710774) Structural visualization/figure preparation/Isolde execution Software, algorithm PyMol Schrödinger Structural visualization/figure preparation Software, algorithm GraphPad Prism 7 GraphPad analysis of signaling data Continued on next page Tsutsumi et al. eLife 2020;9:e58464. .. DOI: https://doi.org/10.7554/eLife.58464 12 of 20 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Other CNBr-Activated Sepharose 4 Fast Flow GE Healthcare 17098101 For preparation of anti-FLAG sepharose Other Lipofectamine 2000 Invitrogen 11668030 Transfection reagent for signaling assay
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