Construct:Article Title: Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected Patients
Article Snippet: .. Mouse CD4 (mCD4)/pVax was constructed by PCR amplification of pBS mCD4 (Addgene, Cambridge, MA) using primers 5 -CCC CCGAATTCCACCATGTGCCGAGCCATCTCTCTT-3 and 5 -CCCCCCTC GAGTCAGATGAGATTATGGCTCTT-3 , which contain the EcoRI and XhoI restriction sites. .. The amplification product was subsequently cloned into the pVAX1 vector (Invitrogen).
Article Title: The herpesviral antagonist m152 reveals differential activation of STING ‐dependent IRF and NF ‐κB signaling and STING 's dual role during MCMV infection
Article Snippet: .. Expression constructs for m152‐V5/His, M35‐V5/His, LacZ‐V5/His, and M27‐V5/His (all in pcDNA3.1‐V5/His, Invitrogen) have been described previously (Munks et al , ). pBS‐mCD4 was purchased from Addgene (#14613) and subcloned into pcDNA3.1‐V5/His via the HindIII / BstXI sites to generate pcDNA3.1 mCD4‐V5/His. pcDNA4 ORF36‐myc/His was previously described (Bussey et al , ) and codes for ORF36 of Kaposi's sarcoma‐associated herpesvirus (KSHV). pPolI A/California/04/2009 NS was kindly provided by Toru Takimoto (University of Rochester Medical Center, USA) and subcloned into pcDNA3.1(‐) via the NotI / EcoRI sites to generate pcDNA3.1(‐) Cal NS1. pEFBOS mCherry‐mSTING (designated Cherry‐STING) expressing monomeric Cherry fused to the N terminus of murine STING and pIRESneo3 cGAS‐GFP (GFP fused to the C terminus of human cGAS) were kindly provided by Andrea Ablasser (Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Switzerland). pcDNA3 hSTING coding for untagged human STING was described previously (Christensen et al , ). pRL‐TK, which expresses Renilla luciferase under control of the thymidine kinase promoter, and pIRES2‐GFP were purchased from Promega and Clontech, respectively. pNF‐κB Luc, containing five NF‐κB responsive elements (TGGGGACTTTCCGC) upstream of the firefly luciferase gene, is commercially available from Agilent Technologies. pGL3basic IFNβ‐Luc (IFNβ‐Luc) and pGL3basic ISG56‐Luc (ISG56‐Luc) were described previously (Chan et al , ). ..
Polymerase Chain Reaction:Article Title: Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected Patients
Article Snippet: .. Mouse CD4 (mCD4)/pVax was constructed by PCR amplification of pBS mCD4 (Addgene, Cambridge, MA) using primers 5 -CCC CCGAATTCCACCATGTGCCGAGCCATCTCTCTT-3 and 5 -CCCCCCTC GAGTCAGATGAGATTATGGCTCTT-3 , which contain the EcoRI and XhoI restriction sites. .. The amplification product was subsequently cloned into the pVAX1 vector (Invitrogen).
Amplification:Article Title: Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected Patients
Article Snippet: .. Mouse CD4 (mCD4)/pVax was constructed by PCR amplification of pBS mCD4 (Addgene, Cambridge, MA) using primers 5 -CCC CCGAATTCCACCATGTGCCGAGCCATCTCTCTT-3 and 5 -CCCCCCTC GAGTCAGATGAGATTATGGCTCTT-3 , which contain the EcoRI and XhoI restriction sites. .. The amplification product was subsequently cloned into the pVAX1 vector (Invitrogen).
Countercurrent Chromatography:Article Title: Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected Patients
Article Snippet: .. Mouse CD4 (mCD4)/pVax was constructed by PCR amplification of pBS mCD4 (Addgene, Cambridge, MA) using primers 5 -CCC CCGAATTCCACCATGTGCCGAGCCATCTCTCTT-3 and 5 -CCCCCCTC GAGTCAGATGAGATTATGGCTCTT-3 , which contain the EcoRI and XhoI restriction sites. .. The amplification product was subsequently cloned into the pVAX1 vector (Invitrogen).
Expressing:Article Title: The herpesviral antagonist m152 reveals differential activation of STING ‐dependent IRF and NF ‐κB signaling and STING 's dual role during MCMV infection
Article Snippet: .. Expression constructs for m152‐V5/His, M35‐V5/His, LacZ‐V5/His, and M27‐V5/His (all in pcDNA3.1‐V5/His, Invitrogen) have been described previously (Munks et al , ). pBS‐mCD4 was purchased from Addgene (#14613) and subcloned into pcDNA3.1‐V5/His via the HindIII / BstXI sites to generate pcDNA3.1 mCD4‐V5/His. pcDNA4 ORF36‐myc/His was previously described (Bussey et al , ) and codes for ORF36 of Kaposi's sarcoma‐associated herpesvirus (KSHV). pPolI A/California/04/2009 NS was kindly provided by Toru Takimoto (University of Rochester Medical Center, USA) and subcloned into pcDNA3.1(‐) via the NotI / EcoRI sites to generate pcDNA3.1(‐) Cal NS1. pEFBOS mCherry‐mSTING (designated Cherry‐STING) expressing monomeric Cherry fused to the N terminus of murine STING and pIRESneo3 cGAS‐GFP (GFP fused to the C terminus of human cGAS) were kindly provided by Andrea Ablasser (Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Switzerland). pcDNA3 hSTING coding for untagged human STING was described previously (Christensen et al , ). pRL‐TK, which expresses Renilla luciferase under control of the thymidine kinase promoter, and pIRES2‐GFP were purchased from Promega and Clontech, respectively. pNF‐κB Luc, containing five NF‐κB responsive elements (TGGGGACTTTCCGC) upstream of the firefly luciferase gene, is commercially available from Agilent Technologies. pGL3basic IFNβ‐Luc (IFNβ‐Luc) and pGL3basic ISG56‐Luc (ISG56‐Luc) were described previously (Chan et al , ). ..
Luciferase:Article Title: The herpesviral antagonist m152 reveals differential activation of STING ‐dependent IRF and NF ‐κB signaling and STING 's dual role during MCMV infection
Article Snippet: .. Expression constructs for m152‐V5/His, M35‐V5/His, LacZ‐V5/His, and M27‐V5/His (all in pcDNA3.1‐V5/His, Invitrogen) have been described previously (Munks et al , ). pBS‐mCD4 was purchased from Addgene (#14613) and subcloned into pcDNA3.1‐V5/His via the HindIII / BstXI sites to generate pcDNA3.1 mCD4‐V5/His. pcDNA4 ORF36‐myc/His was previously described (Bussey et al , ) and codes for ORF36 of Kaposi's sarcoma‐associated herpesvirus (KSHV). pPolI A/California/04/2009 NS was kindly provided by Toru Takimoto (University of Rochester Medical Center, USA) and subcloned into pcDNA3.1(‐) via the NotI / EcoRI sites to generate pcDNA3.1(‐) Cal NS1. pEFBOS mCherry‐mSTING (designated Cherry‐STING) expressing monomeric Cherry fused to the N terminus of murine STING and pIRESneo3 cGAS‐GFP (GFP fused to the C terminus of human cGAS) were kindly provided by Andrea Ablasser (Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Switzerland). pcDNA3 hSTING coding for untagged human STING was described previously (Christensen et al , ). pRL‐TK, which expresses Renilla luciferase under control of the thymidine kinase promoter, and pIRES2‐GFP were purchased from Promega and Clontech, respectively. pNF‐κB Luc, containing five NF‐κB responsive elements (TGGGGACTTTCCGC) upstream of the firefly luciferase gene, is commercially available from Agilent Technologies. pGL3basic IFNβ‐Luc (IFNβ‐Luc) and pGL3basic ISG56‐Luc (ISG56‐Luc) were described previously (Chan et al , ). ..
Control:Article Title: The herpesviral antagonist m152 reveals differential activation of STING ‐dependent IRF and NF ‐κB signaling and STING 's dual role during MCMV infection
Article Snippet: .. Expression constructs for m152‐V5/His, M35‐V5/His, LacZ‐V5/His, and M27‐V5/His (all in pcDNA3.1‐V5/His, Invitrogen) have been described previously (Munks et al , ). pBS‐mCD4 was purchased from Addgene (#14613) and subcloned into pcDNA3.1‐V5/His via the HindIII / BstXI sites to generate pcDNA3.1 mCD4‐V5/His. pcDNA4 ORF36‐myc/His was previously described (Bussey et al , ) and codes for ORF36 of Kaposi's sarcoma‐associated herpesvirus (KSHV). pPolI A/California/04/2009 NS was kindly provided by Toru Takimoto (University of Rochester Medical Center, USA) and subcloned into pcDNA3.1(‐) via the NotI / EcoRI sites to generate pcDNA3.1(‐) Cal NS1. pEFBOS mCherry‐mSTING (designated Cherry‐STING) expressing monomeric Cherry fused to the N terminus of murine STING and pIRESneo3 cGAS‐GFP (GFP fused to the C terminus of human cGAS) were kindly provided by Andrea Ablasser (Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Switzerland). pcDNA3 hSTING coding for untagged human STING was described previously (Christensen et al , ). pRL‐TK, which expresses Renilla luciferase under control of the thymidine kinase promoter, and pIRES2‐GFP were purchased from Promega and Clontech, respectively. pNF‐κB Luc, containing five NF‐κB responsive elements (TGGGGACTTTCCGC) upstream of the firefly luciferase gene, is commercially available from Agilent Technologies. pGL3basic IFNβ‐Luc (IFNβ‐Luc) and pGL3basic ISG56‐Luc (ISG56‐Luc) were described previously (Chan et al , ). ..
Plasmid Preparation:Article Title: Modular Extracellular Sensor Architecture for Engineering Mammalian Cell-based Devices
Article Snippet: .. Source plasmids for MESA components included pCL-CTIG (Addgene plasmid 14901), pRK1043 (Addgene plasmid 8835), pBI-MCS-EGFP (Addgene plasmid 16542), pBS mCD4 (Addgene plasmid 14613), AAV-FLEX-rev-ChR2-tdtomato (Addgene plasmid 18917), pEBFP2-Nuc (Addgene plasmid 14893), YFP-FKBP (Addgene plasmid 20175) and YFP-tagged FRB (YR) (Addgene plasmid 20148), pmCherry-C1 (Clontech 632524). ..
Bioprocessing:Article Title: Modular Extracellular Sensor Architecture for Engineering Mammalian Cell-based Devices
Article Snippet: .. Source plasmids for MESA components included pCL-CTIG (Addgene plasmid 14901), pRK1043 (Addgene plasmid 8835), pBI-MCS-EGFP (Addgene plasmid 16542), pBS mCD4 (Addgene plasmid 14613), AAV-FLEX-rev-ChR2-tdtomato (Addgene plasmid 18917), pEBFP2-Nuc (Addgene plasmid 14893), YFP-FKBP (Addgene plasmid 20175) and YFP-tagged FRB (YR) (Addgene plasmid 20148), pmCherry-C1 (Clontech 632524). ..
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