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    New England Biolabs pcmv gluc
    E2 S266A does not restore repression activity of E8^E2 S78A. RTS3b cells were transfected with 0.5 ng of <t>pCMV-Gluc,</t> 50 ng of pGL31URR reporter plasmid, expression vectors for HPV31 E1 (100 ng), and the indicated expression vectors for HPV31 E2 (10 ng) and E8^E2 (10 ng). Differences in the amounts of DNA were adjusted with the empty expression vectors (pSG5). Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the SEM from four independent experiments performed in duplicate.
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    E2 S266A does not restore repression activity of E8^E2 S78A. RTS3b cells were transfected with 0.5 ng of pCMV-Gluc, 50 ng of pGL31URR reporter plasmid, expression vectors for HPV31 E1 (100 ng), and the indicated expression vectors for HPV31 E2 (10 ng) and E8^E2 (10 ng). Differences in the amounts of DNA were adjusted with the empty expression vectors (pSG5). Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the SEM from four independent experiments performed in duplicate.

    Journal: Journal of Virology

    Article Title: Identification and Functional Characterization of Phosphorylation Sites of the Human Papillomavirus 31 E8^E2 Protein

    doi: 10.1128/JVI.01743-17

    Figure Lengend Snippet: E2 S266A does not restore repression activity of E8^E2 S78A. RTS3b cells were transfected with 0.5 ng of pCMV-Gluc, 50 ng of pGL31URR reporter plasmid, expression vectors for HPV31 E1 (100 ng), and the indicated expression vectors for HPV31 E2 (10 ng) and E8^E2 (10 ng). Differences in the amounts of DNA were adjusted with the empty expression vectors (pSG5). Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the SEM from four independent experiments performed in duplicate.

    Article Snippet: In addition, the pCMV-Gluc plasmid (New England BioLabs) was cotransfected as an internal control.

    Techniques: Activity Assay, Transfection, Plasmid Preparation, Expressing, Luciferase

    Phosphorylation of E8^E2 S78 is required for transcriptional repression of reporter plasmids. (A) HeLa cells were transfected with 100 ng of the pC18-Sp1-luc firefly luciferase reporter, 10 ng of the empty expression vector (pSG5), or the expression vectors for wild-type or mutant HPV31 E8^E2 (left graph) or HPV31 E2 (right graph) and 0.5 ng pCMV-Gluc as an internal control. (B) NHK-HPV31 wild-type cells were transfected with 300 ng pC18-Sp1-luc, 30 ng of the empty vector, or the indicated HPV31 E8^E2 expression vectors (wild type or mutants) and 0.5 ng pCMV-Gluc. Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the standard error of the mean (SEM) from at least seven independent experiments (HeLa) or three independent experiments (NHK-HPV31 WT) performed in duplicate. Statistical significance was determined with a one-way ANOVA and Dunnett's multiple-comparison test: *, P

    Journal: Journal of Virology

    Article Title: Identification and Functional Characterization of Phosphorylation Sites of the Human Papillomavirus 31 E8^E2 Protein

    doi: 10.1128/JVI.01743-17

    Figure Lengend Snippet: Phosphorylation of E8^E2 S78 is required for transcriptional repression of reporter plasmids. (A) HeLa cells were transfected with 100 ng of the pC18-Sp1-luc firefly luciferase reporter, 10 ng of the empty expression vector (pSG5), or the expression vectors for wild-type or mutant HPV31 E8^E2 (left graph) or HPV31 E2 (right graph) and 0.5 ng pCMV-Gluc as an internal control. (B) NHK-HPV31 wild-type cells were transfected with 300 ng pC18-Sp1-luc, 30 ng of the empty vector, or the indicated HPV31 E8^E2 expression vectors (wild type or mutants) and 0.5 ng pCMV-Gluc. Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the standard error of the mean (SEM) from at least seven independent experiments (HeLa) or three independent experiments (NHK-HPV31 WT) performed in duplicate. Statistical significance was determined with a one-way ANOVA and Dunnett's multiple-comparison test: *, P

    Article Snippet: In addition, the pCMV-Gluc plasmid (New England BioLabs) was cotransfected as an internal control.

    Techniques: Transfection, Luciferase, Expressing, Plasmid Preparation, Mutagenesis

    Phosphorylation of E8^E2 S78 is required for repression of replication in a reporter-based replication assay. RTS3b cells were transfected with 0.5 ng of pCMV-Gluc, 50 ng of a reporter plasmid containing the HPV31 URR and the viral early promoter driving the firefly luciferase gene (pGL31URR), and expression vectors for HPV31 E1 (100 ng), E2 (10 ng), and wild-type or mutant E8^E2 (10 ng). Differences in the amounts of DNA were adjusted with the empty expression vectors (pSG5). Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the SEM from five independent experiments performed in duplicate. Statistical significance was determined with a one-way ANOVA and Dunnett's multiple-comparison test: *, P

    Journal: Journal of Virology

    Article Title: Identification and Functional Characterization of Phosphorylation Sites of the Human Papillomavirus 31 E8^E2 Protein

    doi: 10.1128/JVI.01743-17

    Figure Lengend Snippet: Phosphorylation of E8^E2 S78 is required for repression of replication in a reporter-based replication assay. RTS3b cells were transfected with 0.5 ng of pCMV-Gluc, 50 ng of a reporter plasmid containing the HPV31 URR and the viral early promoter driving the firefly luciferase gene (pGL31URR), and expression vectors for HPV31 E1 (100 ng), E2 (10 ng), and wild-type or mutant E8^E2 (10 ng). Differences in the amounts of DNA were adjusted with the empty expression vectors (pSG5). Values are presented as the ratio of firefly luciferase (Fluc) to Gaussia luciferase (Gluc) activities. Error bars indicate the SEM from five independent experiments performed in duplicate. Statistical significance was determined with a one-way ANOVA and Dunnett's multiple-comparison test: *, P

    Article Snippet: In addition, the pCMV-Gluc plasmid (New England BioLabs) was cotransfected as an internal control.

    Techniques: Transfection, Plasmid Preparation, Luciferase, Expressing, Mutagenesis