Recombinant:Article Title: UBE2G1 governs the destruction of cereblon neomorphic substrates
Article Snippet: DOI: https://doi.org/10.7554/eLife.40958 14 of 24 Continued Reagent type Designation Source or reference Identifiers Additional information Recombinant DNA reagent pRSG16-U6sgNT-1-UbiC-TagRFP-2A -Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgCRBN8-UbiC-TagRFP2A-Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2G1-5 -UbiC-TagRFP-2APuro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2 D3-4-UbiC-TagRFP-2A-Puro (plasmid) this paper lentiviral transduction Chemical compound, drug MLN4924 Cayman Cat# 15217 Chemical compound, drug MG-132 R and D systems Cat# 1748/5 Chemical compound, drug cycloheximide EMD Cat# 239765 Chemical compound, drug pomalidomide Celgene CC0004047 Chemical compound, drug lenalidomide Celgene CC0005013 Chemical compound, drug CC-885 Celgene CC0015885 Chemical compound, drug CC-220 Celgene CC0017220 Commercial assay or kit HisPur Ni-NTA Magnetic Beads ThermoFisher Cat# 88832 Peptide, recombinant protein Cul4A-Rbx1 PMID:27338790 Human full length Cul4A and Rbx1 Peptide, recombinant protein Cereblon-DDB1 PMID:27338790 Human cereblon (amino acids 40–442) and full length human DDB1 Peptide, recombinant protein MBP-IKZF1 PMID:27338790 IKZF1 (amino acids 140–168) Peptide, recombinant protein MBP-GSPT1 PMID:27338790 GSPT1 (amino acids 437–633) Peptide, recombinant protein UBE2D1/UbeH5a R and D systems Cat# E2-616-100 Peptide, recombinant protein UBE2D2/UbeH5b R and D systems Cat# E2-622-100 Peptide, recombinant protein UBE2D3/UbeH5c R and D systems Cat# E2-627-100 Peptide, recombinant protein UBE2G1 this paper Human full length UBE2G1 Peptide, recombinant protein wild-type Ubiquitin R and D systems Cat# U-100H Peptide, recombinant protein K48-only Ubiquitin R and D systems Cat# UM-K480-01M Peptide, recombinant protein K48R Ubiquitin R and D systems Cat# UM-K48R-01M Peptide, recombinant protein Ube1 R and D systems Cat# E-305 Lu et al. eLife 2018;7:e40958.
Plasmid Preparation:Article Title: UBE2G1 governs the destruction of cereblon neomorphic substrates
Article Snippet: DOI: https://doi.org/10.7554/eLife.40958 14 of 24 Continued Reagent type Designation Source or reference Identifiers Additional information Recombinant DNA reagent pRSG16-U6sgNT-1-UbiC-TagRFP-2A -Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgCRBN8-UbiC-TagRFP2A-Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2G1-5 -UbiC-TagRFP-2APuro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2 D3-4-UbiC-TagRFP-2A-Puro (plasmid) this paper lentiviral transduction Chemical compound, drug MLN4924 Cayman Cat# 15217 Chemical compound, drug MG-132 R and D systems Cat# 1748/5 Chemical compound, drug cycloheximide EMD Cat# 239765 Chemical compound, drug pomalidomide Celgene CC0004047 Chemical compound, drug lenalidomide Celgene CC0005013 Chemical compound, drug CC-885 Celgene CC0015885 Chemical compound, drug CC-220 Celgene CC0017220 Commercial assay or kit HisPur Ni-NTA Magnetic Beads ThermoFisher Cat# 88832 Peptide, recombinant protein Cul4A-Rbx1 PMID:27338790 Human full length Cul4A and Rbx1 Peptide, recombinant protein Cereblon-DDB1 PMID:27338790 Human cereblon (amino acids 40–442) and full length human DDB1 Peptide, recombinant protein MBP-IKZF1 PMID:27338790 IKZF1 (amino acids 140–168) Peptide, recombinant protein MBP-GSPT1 PMID:27338790 GSPT1 (amino acids 437–633) Peptide, recombinant protein UBE2D1/UbeH5a R and D systems Cat# E2-616-100 Peptide, recombinant protein UBE2D2/UbeH5b R and D systems Cat# E2-622-100 Peptide, recombinant protein UBE2D3/UbeH5c R and D systems Cat# E2-627-100 Peptide, recombinant protein UBE2G1 this paper Human full length UBE2G1 Peptide, recombinant protein wild-type Ubiquitin R and D systems Cat# U-100H Peptide, recombinant protein K48-only Ubiquitin R and D systems Cat# UM-K480-01M Peptide, recombinant protein K48R Ubiquitin R and D systems Cat# UM-K48R-01M Peptide, recombinant protein Ube1 R and D systems Cat# E-305 Lu et al. eLife 2018;7:e40958.
Transduction:Article Title: UBE2G1 governs the destruction of cereblon neomorphic substrates
Article Snippet: DOI: https://doi.org/10.7554/eLife.40958 14 of 24 Continued Reagent type Designation Source or reference Identifiers Additional information Recombinant DNA reagent pRSG16-U6sgNT-1-UbiC-TagRFP-2A -Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgCRBN8-UbiC-TagRFP2A-Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2G1-5 -UbiC-TagRFP-2APuro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2 D3-4-UbiC-TagRFP-2A-Puro (plasmid) this paper lentiviral transduction Chemical compound, drug MLN4924 Cayman Cat# 15217 Chemical compound, drug MG-132 R and D systems Cat# 1748/5 Chemical compound, drug cycloheximide EMD Cat# 239765 Chemical compound, drug pomalidomide Celgene CC0004047 Chemical compound, drug lenalidomide Celgene CC0005013 Chemical compound, drug CC-885 Celgene CC0015885 Chemical compound, drug CC-220 Celgene CC0017220 Commercial assay or kit HisPur Ni-NTA Magnetic Beads ThermoFisher Cat# 88832 Peptide, recombinant protein Cul4A-Rbx1 PMID:27338790 Human full length Cul4A and Rbx1 Peptide, recombinant protein Cereblon-DDB1 PMID:27338790 Human cereblon (amino acids 40–442) and full length human DDB1 Peptide, recombinant protein MBP-IKZF1 PMID:27338790 IKZF1 (amino acids 140–168) Peptide, recombinant protein MBP-GSPT1 PMID:27338790 GSPT1 (amino acids 437–633) Peptide, recombinant protein UBE2D1/UbeH5a R and D systems Cat# E2-616-100 Peptide, recombinant protein UBE2D2/UbeH5b R and D systems Cat# E2-622-100 Peptide, recombinant protein UBE2D3/UbeH5c R and D systems Cat# E2-627-100 Peptide, recombinant protein UBE2G1 this paper Human full length UBE2G1 Peptide, recombinant protein wild-type Ubiquitin R and D systems Cat# U-100H Peptide, recombinant protein K48-only Ubiquitin R and D systems Cat# UM-K480-01M Peptide, recombinant protein K48R Ubiquitin R and D systems Cat# UM-K48R-01M Peptide, recombinant protein Ube1 R and D systems Cat# E-305 Lu et al. eLife 2018;7:e40958.
Magnetic Beads:Article Title: UBE2G1 governs the destruction of cereblon neomorphic substrates
Article Snippet: DOI: https://doi.org/10.7554/eLife.40958 14 of 24 Continued Reagent type Designation Source or reference Identifiers Additional information Recombinant DNA reagent pRSG16-U6sgNT-1-UbiC-TagRFP-2A -Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgCRBN8-UbiC-TagRFP2A-Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2G1-5 -UbiC-TagRFP-2APuro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2 D3-4-UbiC-TagRFP-2A-Puro (plasmid) this paper lentiviral transduction Chemical compound, drug MLN4924 Cayman Cat# 15217 Chemical compound, drug MG-132 R and D systems Cat# 1748/5 Chemical compound, drug cycloheximide EMD Cat# 239765 Chemical compound, drug pomalidomide Celgene CC0004047 Chemical compound, drug lenalidomide Celgene CC0005013 Chemical compound, drug CC-885 Celgene CC0015885 Chemical compound, drug CC-220 Celgene CC0017220 Commercial assay or kit HisPur Ni-NTA Magnetic Beads ThermoFisher Cat# 88832 Peptide, recombinant protein Cul4A-Rbx1 PMID:27338790 Human full length Cul4A and Rbx1 Peptide, recombinant protein Cereblon-DDB1 PMID:27338790 Human cereblon (amino acids 40–442) and full length human DDB1 Peptide, recombinant protein MBP-IKZF1 PMID:27338790 IKZF1 (amino acids 140–168) Peptide, recombinant protein MBP-GSPT1 PMID:27338790 GSPT1 (amino acids 437–633) Peptide, recombinant protein UBE2D1/UbeH5a R and D systems Cat# E2-616-100 Peptide, recombinant protein UBE2D2/UbeH5b R and D systems Cat# E2-622-100 Peptide, recombinant protein UBE2D3/UbeH5c R and D systems Cat# E2-627-100 Peptide, recombinant protein UBE2G1 this paper Human full length UBE2G1 Peptide, recombinant protein wild-type Ubiquitin R and D systems Cat# U-100H Peptide, recombinant protein K48-only Ubiquitin R and D systems Cat# UM-K480-01M Peptide, recombinant protein K48R Ubiquitin R and D systems Cat# UM-K48R-01M Peptide, recombinant protein Ube1 R and D systems Cat# E-305 Lu et al. eLife 2018;7:e40958.
Ubiquitin Proteomics:Article Title: UBE2G1 governs the destruction of cereblon neomorphic substrates
Article Snippet: DOI: https://doi.org/10.7554/eLife.40958 14 of 24 Continued Reagent type Designation Source or reference Identifiers Additional information Recombinant DNA reagent pRSG16-U6sgNT-1-UbiC-TagRFP-2A -Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgCRBN8-UbiC-TagRFP2A-Puro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2G1-5 -UbiC-TagRFP-2APuro (plasmid) this paper lentiviral transduction Recombinant DNA reagent pRSG16-U6-sgUBE2 D3-4-UbiC-TagRFP-2A-Puro (plasmid) this paper lentiviral transduction Chemical compound, drug MLN4924 Cayman Cat# 15217 Chemical compound, drug MG-132 R and D systems Cat# 1748/5 Chemical compound, drug cycloheximide EMD Cat# 239765 Chemical compound, drug pomalidomide Celgene CC0004047 Chemical compound, drug lenalidomide Celgene CC0005013 Chemical compound, drug CC-885 Celgene CC0015885 Chemical compound, drug CC-220 Celgene CC0017220 Commercial assay or kit HisPur Ni-NTA Magnetic Beads ThermoFisher Cat# 88832 Peptide, recombinant protein Cul4A-Rbx1 PMID:27338790 Human full length Cul4A and Rbx1 Peptide, recombinant protein Cereblon-DDB1 PMID:27338790 Human cereblon (amino acids 40–442) and full length human DDB1 Peptide, recombinant protein MBP-IKZF1 PMID:27338790 IKZF1 (amino acids 140–168) Peptide, recombinant protein MBP-GSPT1 PMID:27338790 GSPT1 (amino acids 437–633) Peptide, recombinant protein UBE2D1/UbeH5a R and D systems Cat# E2-616-100 Peptide, recombinant protein UBE2D2/UbeH5b R and D systems Cat# E2-622-100 Peptide, recombinant protein UBE2D3/UbeH5c R and D systems Cat# E2-627-100 Peptide, recombinant protein UBE2G1 this paper Human full length UBE2G1 Peptide, recombinant protein wild-type Ubiquitin R and D systems Cat# U-100H Peptide, recombinant protein K48-only Ubiquitin R and D systems Cat# UM-K480-01M Peptide, recombinant protein K48R Ubiquitin R and D systems Cat# UM-K48R-01M Peptide, recombinant protein Ube1 R and D systems Cat# E-305 Lu et al. eLife 2018;7:e40958.
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