Recombinant:Article Title: An enhanced intracellular delivery platform based on a distant diphtheria toxin homolog that evades pre-existing antitoxin antibodies.
Article Snippet: .. Reagents and tools table Reagent/Resource Reference or Source Identifier or Catalog number Experimental models Vero-nLucP (Cercopithecus aethiops) ATCC, Melnyk Lab (Park et al, 2018) Cat # CCL-81 Hap1 (H. sapiens) Horizon Discovery Cat # C631 Reagent/Resource Reference or Source Identifier or Catalog number HEK293T (H. sapiens) ATCC Cat # CRL-3216 HPAF-II (H. sapiens) ATCC Cat # CRL-1997 MV-4-11 (H. sapiens) ATCC Cat # CRL-9591 SK-N-AS (H. sapiens) ATCC Cat # CRL-2137 Single Donor Human Pediatric Serum age range 4–6 Innovative Research Cat # ISERSPED460 Recombinant DNA ChampionTM pET SUMO Expression System Thermo Fisher Scientific Cat # K30001 lentiCRISPRv2 Addgene Cat # 52961 psPAX2 Addgene Cat # 12260 pMD2.G Addgene Cat # 12259 pLX301 Addgene Cat # 25895 pET-28a EMD Millipore Sigma Cat # 69864 Antibodies Goat Anti-Human IgG+IgM+IgA H&L (HRP), 1/ 10,000 Abcam Cat # ab102420 Oligonucleotides and sequence-based reagents PCR primers This study Table EV1 Chemicals, enzymes and other reagents CloneAmp HiFi PCR Premix Takara Cat # 639298 LB Broth Base (Lennox) Invitrogen by Thermo Fisher Scientific Cat # 12780052 BL21(DE3) Competent E. coli New England Biolabs Cat # C2527H NEB® 5-alpha Competent E. coli (High Efficiency) New England Biolabs Cat # C2987H IPTG Thermo Fisher Scientific Cat # 34060 protease inhibitor cocktail Sigma Aldrich Cat # P8849-5mL PierceTM universal nuclease inhibitor Thermo Fisher Scientific Cat # P88702 Imidazole-HCl Sigma Aldrich Cat # I3386 NaCl Biotech Cat # 7647-14-5 Tris-HCl pH 8.0 Wisent Inc. Cat # 809-128-LL SUMO Protease 1 Life Sensor Cat # SP-4010 EMEM Thermo Fisher Scientific Cat # 11095080 DMEM Thermo Fisher Scientific Cat # 11995073 IMDM Thermo Fisher Scientific Cat # 12440053 © The Author(s) EMBO Molecular Medicine Volume 16 | October 2024 | 2638 – 2651 2645 D ow nloaded from https://w w w .em bopress.org on February 12, 2025 from IP 182.3.51.216. .. Reagent/Resource Reference or Source Identifier or Catalog number MEM NonEssential Amino Acids Solution (100X) Thermo Fisher Scientific Cat # 11140050 FBS, Qualified Thermo Fisher Scientific Cat # 12483-020 PenicillinStreptomycin Thermo Fisher Scientific Cat # 15140122 PrestoBlue Cell Viability Reagent Thermo Fisher Scientific Cat # A13262 SYPRO Orange Invitrogen by Thermo Fisher Scientific Cat # S6651 Tween®-20 Sigma Aldrich Cat # 1379-500mL TBS 10X Wisent Inc. Cat # 311-030-LL TMB Reagent Life Technologies Cat # 002023 Software GraphPad Prism 10.0.0 GraphPad Software https:// www.graphpad.com CFX Manager 3.1 software BioRad https://www.biorad.com/en-ca/sku/ 1845000-cfx-managersoftware?ID=1845000 BLOSUM62 Henikoff and Henikoff (1992) https://doi.org/10.1073/ pnas.89.22.10915 Phenix software package Adams et al (2010) https://www.phenixonline.org/ https://doi.org/10.1107/ S0907444909052925 Coot Emsley et al (2010) https://www2.mrclmb.cam.ac.uk/personal/ pemsley/coot/ https://doi.org/10.1107/ S0907444910007493 Pymol The PyMOL Molecular Graphics System, Version 1.8 Schrödinger, LLC. https://pymol.org/2/ #products Other SpectraMax M5e plate reader Molecular Devices n/a Wizard® Genomic DNA Purification Kit Promega Cat # A1120 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat # G9242 CFX96 qRT-PCR thermocycler BioRad n/a Nano-Glo® Luciferase Assay System Promega Cat # N1130 Presto Mini Plasmid Kit FroggaBio Cat # PDH300 Ni-NTA Magnetic Agarose Thermo Fisher Scientific Cat # 78606 Nunc MaxiSorpTM plates Thermo Fisher Scientific Cat # 12565136 96-well assay plate Corning Cat # CLS3610
Sequencing:Article Title: An enhanced intracellular delivery platform based on a distant diphtheria toxin homolog that evades pre-existing antitoxin antibodies.
Article Snippet: .. Reagents and tools table Reagent/Resource Reference or Source Identifier or Catalog number Experimental models Vero-nLucP (Cercopithecus aethiops) ATCC, Melnyk Lab (Park et al, 2018) Cat # CCL-81 Hap1 (H. sapiens) Horizon Discovery Cat # C631 Reagent/Resource Reference or Source Identifier or Catalog number HEK293T (H. sapiens) ATCC Cat # CRL-3216 HPAF-II (H. sapiens) ATCC Cat # CRL-1997 MV-4-11 (H. sapiens) ATCC Cat # CRL-9591 SK-N-AS (H. sapiens) ATCC Cat # CRL-2137 Single Donor Human Pediatric Serum age range 4–6 Innovative Research Cat # ISERSPED460 Recombinant DNA ChampionTM pET SUMO Expression System Thermo Fisher Scientific Cat # K30001 lentiCRISPRv2 Addgene Cat # 52961 psPAX2 Addgene Cat # 12260 pMD2.G Addgene Cat # 12259 pLX301 Addgene Cat # 25895 pET-28a EMD Millipore Sigma Cat # 69864 Antibodies Goat Anti-Human IgG+IgM+IgA H&L (HRP), 1/ 10,000 Abcam Cat # ab102420 Oligonucleotides and sequence-based reagents PCR primers This study Table EV1 Chemicals, enzymes and other reagents CloneAmp HiFi PCR Premix Takara Cat # 639298 LB Broth Base (Lennox) Invitrogen by Thermo Fisher Scientific Cat # 12780052 BL21(DE3) Competent E. coli New England Biolabs Cat # C2527H NEB® 5-alpha Competent E. coli (High Efficiency) New England Biolabs Cat # C2987H IPTG Thermo Fisher Scientific Cat # 34060 protease inhibitor cocktail Sigma Aldrich Cat # P8849-5mL PierceTM universal nuclease inhibitor Thermo Fisher Scientific Cat # P88702 Imidazole-HCl Sigma Aldrich Cat # I3386 NaCl Biotech Cat # 7647-14-5 Tris-HCl pH 8.0 Wisent Inc. Cat # 809-128-LL SUMO Protease 1 Life Sensor Cat # SP-4010 EMEM Thermo Fisher Scientific Cat # 11095080 DMEM Thermo Fisher Scientific Cat # 11995073 IMDM Thermo Fisher Scientific Cat # 12440053 © The Author(s) EMBO Molecular Medicine Volume 16 | October 2024 | 2638 – 2651 2645 D ow nloaded from https://w w w .em bopress.org on February 12, 2025 from IP 182.3.51.216. .. Reagent/Resource Reference or Source Identifier or Catalog number MEM NonEssential Amino Acids Solution (100X) Thermo Fisher Scientific Cat # 11140050 FBS, Qualified Thermo Fisher Scientific Cat # 12483-020 PenicillinStreptomycin Thermo Fisher Scientific Cat # 15140122 PrestoBlue Cell Viability Reagent Thermo Fisher Scientific Cat # A13262 SYPRO Orange Invitrogen by Thermo Fisher Scientific Cat # S6651 Tween®-20 Sigma Aldrich Cat # 1379-500mL TBS 10X Wisent Inc. Cat # 311-030-LL TMB Reagent Life Technologies Cat # 002023 Software GraphPad Prism 10.0.0 GraphPad Software https:// www.graphpad.com CFX Manager 3.1 software BioRad https://www.biorad.com/en-ca/sku/ 1845000-cfx-managersoftware?ID=1845000 BLOSUM62 Henikoff and Henikoff (1992) https://doi.org/10.1073/ pnas.89.22.10915 Phenix software package Adams et al (2010) https://www.phenixonline.org/ https://doi.org/10.1107/ S0907444909052925 Coot Emsley et al (2010) https://www2.mrclmb.cam.ac.uk/personal/ pemsley/coot/ https://doi.org/10.1107/ S0907444910007493 Pymol The PyMOL Molecular Graphics System, Version 1.8 Schrödinger, LLC. https://pymol.org/2/ #products Other SpectraMax M5e plate reader Molecular Devices n/a Wizard® Genomic DNA Purification Kit Promega Cat # A1120 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat # G9242 CFX96 qRT-PCR thermocycler BioRad n/a Nano-Glo® Luciferase Assay System Promega Cat # N1130 Presto Mini Plasmid Kit FroggaBio Cat # PDH300 Ni-NTA Magnetic Agarose Thermo Fisher Scientific Cat # 78606 Nunc MaxiSorpTM plates Thermo Fisher Scientific Cat # 12565136 96-well assay plate Corning Cat # CLS3610
Polymerase Chain Reaction:Article Title: An enhanced intracellular delivery platform based on a distant diphtheria toxin homolog that evades pre-existing antitoxin antibodies.
Article Snippet: .. Reagents and tools table Reagent/Resource Reference or Source Identifier or Catalog number Experimental models Vero-nLucP (Cercopithecus aethiops) ATCC, Melnyk Lab (Park et al, 2018) Cat # CCL-81 Hap1 (H. sapiens) Horizon Discovery Cat # C631 Reagent/Resource Reference or Source Identifier or Catalog number HEK293T (H. sapiens) ATCC Cat # CRL-3216 HPAF-II (H. sapiens) ATCC Cat # CRL-1997 MV-4-11 (H. sapiens) ATCC Cat # CRL-9591 SK-N-AS (H. sapiens) ATCC Cat # CRL-2137 Single Donor Human Pediatric Serum age range 4–6 Innovative Research Cat # ISERSPED460 Recombinant DNA ChampionTM pET SUMO Expression System Thermo Fisher Scientific Cat # K30001 lentiCRISPRv2 Addgene Cat # 52961 psPAX2 Addgene Cat # 12260 pMD2.G Addgene Cat # 12259 pLX301 Addgene Cat # 25895 pET-28a EMD Millipore Sigma Cat # 69864 Antibodies Goat Anti-Human IgG+IgM+IgA H&L (HRP), 1/ 10,000 Abcam Cat # ab102420 Oligonucleotides and sequence-based reagents PCR primers This study Table EV1 Chemicals, enzymes and other reagents CloneAmp HiFi PCR Premix Takara Cat # 639298 LB Broth Base (Lennox) Invitrogen by Thermo Fisher Scientific Cat # 12780052 BL21(DE3) Competent E. coli New England Biolabs Cat # C2527H NEB® 5-alpha Competent E. coli (High Efficiency) New England Biolabs Cat # C2987H IPTG Thermo Fisher Scientific Cat # 34060 protease inhibitor cocktail Sigma Aldrich Cat # P8849-5mL PierceTM universal nuclease inhibitor Thermo Fisher Scientific Cat # P88702 Imidazole-HCl Sigma Aldrich Cat # I3386 NaCl Biotech Cat # 7647-14-5 Tris-HCl pH 8.0 Wisent Inc. Cat # 809-128-LL SUMO Protease 1 Life Sensor Cat # SP-4010 EMEM Thermo Fisher Scientific Cat # 11095080 DMEM Thermo Fisher Scientific Cat # 11995073 IMDM Thermo Fisher Scientific Cat # 12440053 © The Author(s) EMBO Molecular Medicine Volume 16 | October 2024 | 2638 – 2651 2645 D ow nloaded from https://w w w .em bopress.org on February 12, 2025 from IP 182.3.51.216. .. Reagent/Resource Reference or Source Identifier or Catalog number MEM NonEssential Amino Acids Solution (100X) Thermo Fisher Scientific Cat # 11140050 FBS, Qualified Thermo Fisher Scientific Cat # 12483-020 PenicillinStreptomycin Thermo Fisher Scientific Cat # 15140122 PrestoBlue Cell Viability Reagent Thermo Fisher Scientific Cat # A13262 SYPRO Orange Invitrogen by Thermo Fisher Scientific Cat # S6651 Tween®-20 Sigma Aldrich Cat # 1379-500mL TBS 10X Wisent Inc. Cat # 311-030-LL TMB Reagent Life Technologies Cat # 002023 Software GraphPad Prism 10.0.0 GraphPad Software https:// www.graphpad.com CFX Manager 3.1 software BioRad https://www.biorad.com/en-ca/sku/ 1845000-cfx-managersoftware?ID=1845000 BLOSUM62 Henikoff and Henikoff (1992) https://doi.org/10.1073/ pnas.89.22.10915 Phenix software package Adams et al (2010) https://www.phenixonline.org/ https://doi.org/10.1107/ S0907444909052925 Coot Emsley et al (2010) https://www2.mrclmb.cam.ac.uk/personal/ pemsley/coot/ https://doi.org/10.1107/ S0907444910007493 Pymol The PyMOL Molecular Graphics System, Version 1.8 Schrödinger, LLC. https://pymol.org/2/ #products Other SpectraMax M5e plate reader Molecular Devices n/a Wizard® Genomic DNA Purification Kit Promega Cat # A1120 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat # G9242 CFX96 qRT-PCR thermocycler BioRad n/a Nano-Glo® Luciferase Assay System Promega Cat # N1130 Presto Mini Plasmid Kit FroggaBio Cat # PDH300 Ni-NTA Magnetic Agarose Thermo Fisher Scientific Cat # 78606 Nunc MaxiSorpTM plates Thermo Fisher Scientific Cat # 12565136 96-well assay plate Corning Cat # CLS3610
Protease Inhibitor:Article Title: An enhanced intracellular delivery platform based on a distant diphtheria toxin homolog that evades pre-existing antitoxin antibodies.
Article Snippet: .. Reagents and tools table Reagent/Resource Reference or Source Identifier or Catalog number Experimental models Vero-nLucP (Cercopithecus aethiops) ATCC, Melnyk Lab (Park et al, 2018) Cat # CCL-81 Hap1 (H. sapiens) Horizon Discovery Cat # C631 Reagent/Resource Reference or Source Identifier or Catalog number HEK293T (H. sapiens) ATCC Cat # CRL-3216 HPAF-II (H. sapiens) ATCC Cat # CRL-1997 MV-4-11 (H. sapiens) ATCC Cat # CRL-9591 SK-N-AS (H. sapiens) ATCC Cat # CRL-2137 Single Donor Human Pediatric Serum age range 4–6 Innovative Research Cat # ISERSPED460 Recombinant DNA ChampionTM pET SUMO Expression System Thermo Fisher Scientific Cat # K30001 lentiCRISPRv2 Addgene Cat # 52961 psPAX2 Addgene Cat # 12260 pMD2.G Addgene Cat # 12259 pLX301 Addgene Cat # 25895 pET-28a EMD Millipore Sigma Cat # 69864 Antibodies Goat Anti-Human IgG+IgM+IgA H&L (HRP), 1/ 10,000 Abcam Cat # ab102420 Oligonucleotides and sequence-based reagents PCR primers This study Table EV1 Chemicals, enzymes and other reagents CloneAmp HiFi PCR Premix Takara Cat # 639298 LB Broth Base (Lennox) Invitrogen by Thermo Fisher Scientific Cat # 12780052 BL21(DE3) Competent E. coli New England Biolabs Cat # C2527H NEB® 5-alpha Competent E. coli (High Efficiency) New England Biolabs Cat # C2987H IPTG Thermo Fisher Scientific Cat # 34060 protease inhibitor cocktail Sigma Aldrich Cat # P8849-5mL PierceTM universal nuclease inhibitor Thermo Fisher Scientific Cat # P88702 Imidazole-HCl Sigma Aldrich Cat # I3386 NaCl Biotech Cat # 7647-14-5 Tris-HCl pH 8.0 Wisent Inc. Cat # 809-128-LL SUMO Protease 1 Life Sensor Cat # SP-4010 EMEM Thermo Fisher Scientific Cat # 11095080 DMEM Thermo Fisher Scientific Cat # 11995073 IMDM Thermo Fisher Scientific Cat # 12440053 © The Author(s) EMBO Molecular Medicine Volume 16 | October 2024 | 2638 – 2651 2645 D ow nloaded from https://w w w .em bopress.org on February 12, 2025 from IP 182.3.51.216. .. Reagent/Resource Reference or Source Identifier or Catalog number MEM NonEssential Amino Acids Solution (100X) Thermo Fisher Scientific Cat # 11140050 FBS, Qualified Thermo Fisher Scientific Cat # 12483-020 PenicillinStreptomycin Thermo Fisher Scientific Cat # 15140122 PrestoBlue Cell Viability Reagent Thermo Fisher Scientific Cat # A13262 SYPRO Orange Invitrogen by Thermo Fisher Scientific Cat # S6651 Tween®-20 Sigma Aldrich Cat # 1379-500mL TBS 10X Wisent Inc. Cat # 311-030-LL TMB Reagent Life Technologies Cat # 002023 Software GraphPad Prism 10.0.0 GraphPad Software https:// www.graphpad.com CFX Manager 3.1 software BioRad https://www.biorad.com/en-ca/sku/ 1845000-cfx-managersoftware?ID=1845000 BLOSUM62 Henikoff and Henikoff (1992) https://doi.org/10.1073/ pnas.89.22.10915 Phenix software package Adams et al (2010) https://www.phenixonline.org/ https://doi.org/10.1107/ S0907444909052925 Coot Emsley et al (2010) https://www2.mrclmb.cam.ac.uk/personal/ pemsley/coot/ https://doi.org/10.1107/ S0907444910007493 Pymol The PyMOL Molecular Graphics System, Version 1.8 Schrödinger, LLC. https://pymol.org/2/ #products Other SpectraMax M5e plate reader Molecular Devices n/a Wizard® Genomic DNA Purification Kit Promega Cat # A1120 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat # G9242 CFX96 qRT-PCR thermocycler BioRad n/a Nano-Glo® Luciferase Assay System Promega Cat # N1130 Presto Mini Plasmid Kit FroggaBio Cat # PDH300 Ni-NTA Magnetic Agarose Thermo Fisher Scientific Cat # 78606 Nunc MaxiSorpTM plates Thermo Fisher Scientific Cat # 12565136 96-well assay plate Corning Cat # CLS3610
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