Produced:Article Title: Chaperone Function of Hgh1 in the Biogenesis of Eukaryotic Elongation Factor 2.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Rabbit IgG (whole molecule)-Peroxidase antibody produced in goat Sigma-Aldrich Cat. #A6154 Goat anti-mouse IgG HRP conjugated Dako Cat. #P0447 Mouse monoclonal anti-FLAG Sigma-Aldrich Cat. #F3165 Mouse monoclonal anti-Pgk1 Life Technologies Cat. #459250 Rabbit anti-Hgh1 antiserum This study N/A Rabbit anti-Tcp1 (Behrends et al., 2006) N/A Rabbit polyclonal anti-eEF2 Aviva Cat. #ARP58457_P050 Rat monoclonal anti-HA High Affinity, 3F10 Merck Cat. #ROAHAHA Bacterial and Virus Strains Escherichia coli Bl21 codon+ RIL Agilent Cat. #NC9122855 Chemicals, Peptides, and Recombinant Proteins Benzonase Novagen Cat. #707416 Complete protease inhibitor cocktail, EDTA-free Roche Cat. #05056489001 Cycloheximide Sigma-Aldrich Cat. #C4859 Disuccinimidyl suberate (DSS) d0/d12 Creative Molecules Cat. #001S EheI (SfoI) restriction enzyme Thermo Fisher Scientific Cat. #ER0441 FLAG peptide Sigma-Aldrich Cat. #F3290 Geneticin selective antibiotic (G418 sulfate) Thermo Fisher Scientific Cat. #11811031 GMP-PNP Na salt Jena Bioscience Cat. #NU-401-50 Hygromycin B Calbiochem Cat. #400051 Kod hot start DNA polymerase Novagen Cat. #71086-3 Macbecin I Tocris Cat. #3061 NuPAGE 3–8% Tris-Acetate Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #EA0375BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #NP0321BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 15-well Invitrogen Cat. #NP0323BOX Pfu DNA polymerase Promega Cat. # M7745 Puromycin dihydrochloride GIBCO BRL Cat. #A11138 Sordarin Sigma-Aldrich Cat. #S1442 T4 DNA ligase New England Biolabs Cat. # M0202S TCEP Thermo Fisher Scientific Cat. #20491 Critical Commercial Assays Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat. #500-0006 Luminata Classico Merck Cat. #WBLUC0500 Deposited Data Coordinates and structure factors: Hgh1(1-363) Se-SAD This study PDB: 6HBB Coordinates and structure factors: Hgh1(1-363), crystal form I This study PDB: 6HBA Coordinates and structure factors: Hgh1(1-363), crystal form II This study PDB: 6HBC Immunoblot data: (Mendeley Data) This study https://doi.org/10.17632/sw9sxts73z.1 Experimental Models: Organisms/Strains BY4741 MATa his3D1 leu2D0 lys2D0 ura3D0 EUROSCARF http://www.euroscarf.de BY4741 hgh1D::KanMX4 This study yCLK14 BY4741 eft2D::URA3 This study yCLK17 (Continued on next page) e1 Molecular Cell 74, 1–13.e1–e9, April 4, 2019
Virus:Article Title: Chaperone Function of Hgh1 in the Biogenesis of Eukaryotic Elongation Factor 2.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Rabbit IgG (whole molecule)-Peroxidase antibody produced in goat Sigma-Aldrich Cat. #A6154 Goat anti-mouse IgG HRP conjugated Dako Cat. #P0447 Mouse monoclonal anti-FLAG Sigma-Aldrich Cat. #F3165 Mouse monoclonal anti-Pgk1 Life Technologies Cat. #459250 Rabbit anti-Hgh1 antiserum This study N/A Rabbit anti-Tcp1 (Behrends et al., 2006) N/A Rabbit polyclonal anti-eEF2 Aviva Cat. #ARP58457_P050 Rat monoclonal anti-HA High Affinity, 3F10 Merck Cat. #ROAHAHA Bacterial and Virus Strains Escherichia coli Bl21 codon+ RIL Agilent Cat. #NC9122855 Chemicals, Peptides, and Recombinant Proteins Benzonase Novagen Cat. #707416 Complete protease inhibitor cocktail, EDTA-free Roche Cat. #05056489001 Cycloheximide Sigma-Aldrich Cat. #C4859 Disuccinimidyl suberate (DSS) d0/d12 Creative Molecules Cat. #001S EheI (SfoI) restriction enzyme Thermo Fisher Scientific Cat. #ER0441 FLAG peptide Sigma-Aldrich Cat. #F3290 Geneticin selective antibiotic (G418 sulfate) Thermo Fisher Scientific Cat. #11811031 GMP-PNP Na salt Jena Bioscience Cat. #NU-401-50 Hygromycin B Calbiochem Cat. #400051 Kod hot start DNA polymerase Novagen Cat. #71086-3 Macbecin I Tocris Cat. #3061 NuPAGE 3–8% Tris-Acetate Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #EA0375BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #NP0321BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 15-well Invitrogen Cat. #NP0323BOX Pfu DNA polymerase Promega Cat. # M7745 Puromycin dihydrochloride GIBCO BRL Cat. #A11138 Sordarin Sigma-Aldrich Cat. #S1442 T4 DNA ligase New England Biolabs Cat. # M0202S TCEP Thermo Fisher Scientific Cat. #20491 Critical Commercial Assays Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat. #500-0006 Luminata Classico Merck Cat. #WBLUC0500 Deposited Data Coordinates and structure factors: Hgh1(1-363) Se-SAD This study PDB: 6HBB Coordinates and structure factors: Hgh1(1-363), crystal form I This study PDB: 6HBA Coordinates and structure factors: Hgh1(1-363), crystal form II This study PDB: 6HBC Immunoblot data: (Mendeley Data) This study https://doi.org/10.17632/sw9sxts73z.1 Experimental Models: Organisms/Strains BY4741 MATa his3D1 leu2D0 lys2D0 ura3D0 EUROSCARF http://www.euroscarf.de BY4741 hgh1D::KanMX4 This study yCLK14 BY4741 eft2D::URA3 This study yCLK17 (Continued on next page) e1 Molecular Cell 74, 1–13.e1–e9, April 4, 2019
Recombinant:Article Title: Chaperone Function of Hgh1 in the Biogenesis of Eukaryotic Elongation Factor 2.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Rabbit IgG (whole molecule)-Peroxidase antibody produced in goat Sigma-Aldrich Cat. #A6154 Goat anti-mouse IgG HRP conjugated Dako Cat. #P0447 Mouse monoclonal anti-FLAG Sigma-Aldrich Cat. #F3165 Mouse monoclonal anti-Pgk1 Life Technologies Cat. #459250 Rabbit anti-Hgh1 antiserum This study N/A Rabbit anti-Tcp1 (Behrends et al., 2006) N/A Rabbit polyclonal anti-eEF2 Aviva Cat. #ARP58457_P050 Rat monoclonal anti-HA High Affinity, 3F10 Merck Cat. #ROAHAHA Bacterial and Virus Strains Escherichia coli Bl21 codon+ RIL Agilent Cat. #NC9122855 Chemicals, Peptides, and Recombinant Proteins Benzonase Novagen Cat. #707416 Complete protease inhibitor cocktail, EDTA-free Roche Cat. #05056489001 Cycloheximide Sigma-Aldrich Cat. #C4859 Disuccinimidyl suberate (DSS) d0/d12 Creative Molecules Cat. #001S EheI (SfoI) restriction enzyme Thermo Fisher Scientific Cat. #ER0441 FLAG peptide Sigma-Aldrich Cat. #F3290 Geneticin selective antibiotic (G418 sulfate) Thermo Fisher Scientific Cat. #11811031 GMP-PNP Na salt Jena Bioscience Cat. #NU-401-50 Hygromycin B Calbiochem Cat. #400051 Kod hot start DNA polymerase Novagen Cat. #71086-3 Macbecin I Tocris Cat. #3061 NuPAGE 3–8% Tris-Acetate Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #EA0375BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #NP0321BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 15-well Invitrogen Cat. #NP0323BOX Pfu DNA polymerase Promega Cat. # M7745 Puromycin dihydrochloride GIBCO BRL Cat. #A11138 Sordarin Sigma-Aldrich Cat. #S1442 T4 DNA ligase New England Biolabs Cat. # M0202S TCEP Thermo Fisher Scientific Cat. #20491 Critical Commercial Assays Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat. #500-0006 Luminata Classico Merck Cat. #WBLUC0500 Deposited Data Coordinates and structure factors: Hgh1(1-363) Se-SAD This study PDB: 6HBB Coordinates and structure factors: Hgh1(1-363), crystal form I This study PDB: 6HBA Coordinates and structure factors: Hgh1(1-363), crystal form II This study PDB: 6HBC Immunoblot data: (Mendeley Data) This study https://doi.org/10.17632/sw9sxts73z.1 Experimental Models: Organisms/Strains BY4741 MATa his3D1 leu2D0 lys2D0 ura3D0 EUROSCARF http://www.euroscarf.de BY4741 hgh1D::KanMX4 This study yCLK14 BY4741 eft2D::URA3 This study yCLK17 (Continued on next page) e1 Molecular Cell 74, 1–13.e1–e9, April 4, 2019
Protease Inhibitor:Article Title: Chaperone Function of Hgh1 in the Biogenesis of Eukaryotic Elongation Factor 2.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Rabbit IgG (whole molecule)-Peroxidase antibody produced in goat Sigma-Aldrich Cat. #A6154 Goat anti-mouse IgG HRP conjugated Dako Cat. #P0447 Mouse monoclonal anti-FLAG Sigma-Aldrich Cat. #F3165 Mouse monoclonal anti-Pgk1 Life Technologies Cat. #459250 Rabbit anti-Hgh1 antiserum This study N/A Rabbit anti-Tcp1 (Behrends et al., 2006) N/A Rabbit polyclonal anti-eEF2 Aviva Cat. #ARP58457_P050 Rat monoclonal anti-HA High Affinity, 3F10 Merck Cat. #ROAHAHA Bacterial and Virus Strains Escherichia coli Bl21 codon+ RIL Agilent Cat. #NC9122855 Chemicals, Peptides, and Recombinant Proteins Benzonase Novagen Cat. #707416 Complete protease inhibitor cocktail, EDTA-free Roche Cat. #05056489001 Cycloheximide Sigma-Aldrich Cat. #C4859 Disuccinimidyl suberate (DSS) d0/d12 Creative Molecules Cat. #001S EheI (SfoI) restriction enzyme Thermo Fisher Scientific Cat. #ER0441 FLAG peptide Sigma-Aldrich Cat. #F3290 Geneticin selective antibiotic (G418 sulfate) Thermo Fisher Scientific Cat. #11811031 GMP-PNP Na salt Jena Bioscience Cat. #NU-401-50 Hygromycin B Calbiochem Cat. #400051 Kod hot start DNA polymerase Novagen Cat. #71086-3 Macbecin I Tocris Cat. #3061 NuPAGE 3–8% Tris-Acetate Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #EA0375BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #NP0321BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 15-well Invitrogen Cat. #NP0323BOX Pfu DNA polymerase Promega Cat. # M7745 Puromycin dihydrochloride GIBCO BRL Cat. #A11138 Sordarin Sigma-Aldrich Cat. #S1442 T4 DNA ligase New England Biolabs Cat. # M0202S TCEP Thermo Fisher Scientific Cat. #20491 Critical Commercial Assays Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat. #500-0006 Luminata Classico Merck Cat. #WBLUC0500 Deposited Data Coordinates and structure factors: Hgh1(1-363) Se-SAD This study PDB: 6HBB Coordinates and structure factors: Hgh1(1-363), crystal form I This study PDB: 6HBA Coordinates and structure factors: Hgh1(1-363), crystal form II This study PDB: 6HBC Immunoblot data: (Mendeley Data) This study https://doi.org/10.17632/sw9sxts73z.1 Experimental Models: Organisms/Strains BY4741 MATa his3D1 leu2D0 lys2D0 ura3D0 EUROSCARF http://www.euroscarf.de BY4741 hgh1D::KanMX4 This study yCLK14 BY4741 eft2D::URA3 This study yCLK17 (Continued on next page) e1 Molecular Cell 74, 1–13.e1–e9, April 4, 2019
Western Blot:Article Title: Chaperone Function of Hgh1 in the Biogenesis of Eukaryotic Elongation Factor 2.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Rabbit IgG (whole molecule)-Peroxidase antibody produced in goat Sigma-Aldrich Cat. #A6154 Goat anti-mouse IgG HRP conjugated Dako Cat. #P0447 Mouse monoclonal anti-FLAG Sigma-Aldrich Cat. #F3165 Mouse monoclonal anti-Pgk1 Life Technologies Cat. #459250 Rabbit anti-Hgh1 antiserum This study N/A Rabbit anti-Tcp1 (Behrends et al., 2006) N/A Rabbit polyclonal anti-eEF2 Aviva Cat. #ARP58457_P050 Rat monoclonal anti-HA High Affinity, 3F10 Merck Cat. #ROAHAHA Bacterial and Virus Strains Escherichia coli Bl21 codon+ RIL Agilent Cat. #NC9122855 Chemicals, Peptides, and Recombinant Proteins Benzonase Novagen Cat. #707416 Complete protease inhibitor cocktail, EDTA-free Roche Cat. #05056489001 Cycloheximide Sigma-Aldrich Cat. #C4859 Disuccinimidyl suberate (DSS) d0/d12 Creative Molecules Cat. #001S EheI (SfoI) restriction enzyme Thermo Fisher Scientific Cat. #ER0441 FLAG peptide Sigma-Aldrich Cat. #F3290 Geneticin selective antibiotic (G418 sulfate) Thermo Fisher Scientific Cat. #11811031 GMP-PNP Na salt Jena Bioscience Cat. #NU-401-50 Hygromycin B Calbiochem Cat. #400051 Kod hot start DNA polymerase Novagen Cat. #71086-3 Macbecin I Tocris Cat. #3061 NuPAGE 3–8% Tris-Acetate Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #EA0375BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 10-well Invitrogen Cat. #NP0321BOX NuPAGE 4–12% Bis-Tris Protein Gels, 1.0 mm, 15-well Invitrogen Cat. #NP0323BOX Pfu DNA polymerase Promega Cat. # M7745 Puromycin dihydrochloride GIBCO BRL Cat. #A11138 Sordarin Sigma-Aldrich Cat. #S1442 T4 DNA ligase New England Biolabs Cat. # M0202S TCEP Thermo Fisher Scientific Cat. #20491 Critical Commercial Assays Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat. #500-0006 Luminata Classico Merck Cat. #WBLUC0500 Deposited Data Coordinates and structure factors: Hgh1(1-363) Se-SAD This study PDB: 6HBB Coordinates and structure factors: Hgh1(1-363), crystal form I This study PDB: 6HBA Coordinates and structure factors: Hgh1(1-363), crystal form II This study PDB: 6HBC Immunoblot data: (Mendeley Data) This study https://doi.org/10.17632/sw9sxts73z.1 Experimental Models: Organisms/Strains BY4741 MATa his3D1 leu2D0 lys2D0 ura3D0 EUROSCARF http://www.euroscarf.de BY4741 hgh1D::KanMX4 This study yCLK14 BY4741 eft2D::URA3 This study yCLK17 (Continued on next page) e1 Molecular Cell 74, 1–13.e1–e9, April 4, 2019
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