dual-luciferase reporter assay system Search Results


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  • 88
    Promega dual luc reporter assay system
    Dual Luc Reporter Assay System, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 132 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega dual luciferase reporter gene activity assay a dual luciferase reporter assay system kit
    Dual Luciferase Reporter Gene Activity Assay A Dual Luciferase Reporter Assay System Kit, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega luciferase activity assay a dual luciferase reporter system
    Luciferase Activity Assay A Dual Luciferase Reporter System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dual luciferase reporter assay system
    PRL-3 transcriptionally regulates the expression of RAP1. Notes: ( A ) Western blot results showed that overexpression of PRL-3 increased the RAP1 level (left), and silencing of PRL-3 decreased RAP1 level (right). ( B ) RT-PCR results showed that overexpression of PRL-3 increased RAP1 level (upper) and silencing of PRL-3 decreased RAP1 level (lower). ( C ) <t>Dual</t> <t>luciferase</t> <t>reporter</t> assays of rap1 gene promoter in HCT116 and 293 T cells expressing pcDNA3.0-myc-NC/PRL-3 and pGLB-RAP1(−1,500/+150)/(−500/+150). Results were normalized to the activity obtained in cells transfected with pcDNA3.0-myc-NC and pGLB-RAP1 (−1,500/+150). ( D ) ChIP <t>assay</t> using PRL-3 or IgG in HCT116 cells, and qPCR of different promoter regions of rap1 gene. Values correspond to the ratio between the PRL-3 antibody immunoprecipitated DNA with respect to the IgG immunoprecipitated DNA. ( E ) Gene expression correlation analysis between PRL-3 and RAP1 in TCGA data ( P =0.0005, R =0.52) and GTEx data ( P
    Dual Luciferase Reporter Assay System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Guangzhou RiboBio dual luciferase reporter assay system
    PRL-3 transcriptionally regulates the expression of RAP1. Notes: ( A ) Western blot results showed that overexpression of PRL-3 increased the RAP1 level (left), and silencing of PRL-3 decreased RAP1 level (right). ( B ) RT-PCR results showed that overexpression of PRL-3 increased RAP1 level (upper) and silencing of PRL-3 decreased RAP1 level (lower). ( C ) <t>Dual</t> <t>luciferase</t> <t>reporter</t> assays of rap1 gene promoter in HCT116 and 293 T cells expressing pcDNA3.0-myc-NC/PRL-3 and pGLB-RAP1(−1,500/+150)/(−500/+150). Results were normalized to the activity obtained in cells transfected with pcDNA3.0-myc-NC and pGLB-RAP1 (−1,500/+150). ( D ) ChIP <t>assay</t> using PRL-3 or IgG in HCT116 cells, and qPCR of different promoter regions of rap1 gene. Values correspond to the ratio between the PRL-3 antibody immunoprecipitated DNA with respect to the IgG immunoprecipitated DNA. ( E ) Gene expression correlation analysis between PRL-3 and RAP1 in TCGA data ( P =0.0005, R =0.52) and GTEx data ( P
    Dual Luciferase Reporter Assay System, supplied by Guangzhou RiboBio, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ribobio dual luciferase reporter assay system
    PRL-3 transcriptionally regulates the expression of RAP1. Notes: ( A ) Western blot results showed that overexpression of PRL-3 increased the RAP1 level (left), and silencing of PRL-3 decreased RAP1 level (right). ( B ) RT-PCR results showed that overexpression of PRL-3 increased RAP1 level (upper) and silencing of PRL-3 decreased RAP1 level (lower). ( C ) <t>Dual</t> <t>luciferase</t> <t>reporter</t> assays of rap1 gene promoter in HCT116 and 293 T cells expressing pcDNA3.0-myc-NC/PRL-3 and pGLB-RAP1(−1,500/+150)/(−500/+150). Results were normalized to the activity obtained in cells transfected with pcDNA3.0-myc-NC and pGLB-RAP1 (−1,500/+150). ( D ) ChIP <t>assay</t> using PRL-3 or IgG in HCT116 cells, and qPCR of different promoter regions of rap1 gene. Values correspond to the ratio between the PRL-3 antibody immunoprecipitated DNA with respect to the IgG immunoprecipitated DNA. ( E ) Gene expression correlation analysis between PRL-3 and RAP1 in TCGA data ( P =0.0005, R =0.52) and GTEx data ( P
    Dual Luciferase Reporter Assay System, supplied by ribobio, used in various techniques. Bioz Stars score: 91/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Beyotime dual luciferase reporter assay system
    PDGFRB is a direct target gene of miR-518b. (A) Inverse relationship between the expression of miR-518b and PDGFRB was detected in GBM specimens (magnification, 200x). (B) Western blot analysis of PDGFRB expression in U87 and U251 cells 48 h post-transfection with miR-518b and miR-ctrl. (C) PDGFRB mRNA 3′UTR contains binding sites for miR-518b. (D) U87 and U251 cells were co-transfected with the <t>dual-luciferase</t> <t>reporter</t> plasmid carrying the Wt or Mut 3′UTR sequences of PDGFRB and miR-518b or miR-ctrl mimics. A luciferase reporter <t>system</t> analysis was performed. Data are presented as the mean ± standard deviation (n=5). *P
    Dual Luciferase Reporter Assay System, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 288 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Keygen Biotech dual luciferase reporter assay system
    PDGFRB is a direct target gene of miR-518b. (A) Inverse relationship between the expression of miR-518b and PDGFRB was detected in GBM specimens (magnification, 200x). (B) Western blot analysis of PDGFRB expression in U87 and U251 cells 48 h post-transfection with miR-518b and miR-ctrl. (C) PDGFRB mRNA 3′UTR contains binding sites for miR-518b. (D) U87 and U251 cells were co-transfected with the <t>dual-luciferase</t> <t>reporter</t> plasmid carrying the Wt or Mut 3′UTR sequences of PDGFRB and miR-518b or miR-ctrl mimics. A luciferase reporter <t>system</t> analysis was performed. Data are presented as the mean ± standard deviation (n=5). *P
    Dual Luciferase Reporter Assay System, supplied by Keygen Biotech, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega dual luciferase reporter 1000 assay systems
    PDGFRB is a direct target gene of miR-518b. (A) Inverse relationship between the expression of miR-518b and PDGFRB was detected in GBM specimens (magnification, 200x). (B) Western blot analysis of PDGFRB expression in U87 and U251 cells 48 h post-transfection with miR-518b and miR-ctrl. (C) PDGFRB mRNA 3′UTR contains binding sites for miR-518b. (D) U87 and U251 cells were co-transfected with the <t>dual-luciferase</t> <t>reporter</t> plasmid carrying the Wt or Mut 3′UTR sequences of PDGFRB and miR-518b or miR-ctrl mimics. A luciferase reporter <t>system</t> analysis was performed. Data are presented as the mean ± standard deviation (n=5). *P
    Dual Luciferase Reporter 1000 Assay Systems, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TransGen biotech co dual luciferase reporter assay system
    miR-155 directly targeted on MafB. (a) The target site of miR-155 and MafB 3′UTR was measured based on TargetScan and microRNA.org. (b) Investigation of the regulatory effect of miR-155 on MafB expression using <t>dual-luciferase</t> <t>reporter</t> <t>assay</t> in 293T cells. (c) and (d and e) The expression of MafB in OGD/R-induced SH-SY5Y cells transfected with miR-155 or anti-miR-155 was measured through qRT-PCR and western blot analyses respectively ∗ P
    Dual Luciferase Reporter Assay System, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genecopoeia dual luciferase reporter assay system
    HOTAIR modulates DNA methylation within MTHFR promoter region. shRNA was delivered into TE-1 cells to specifically blunt HOTAIR (sh-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1 cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells were served as negative controls (sh-NC group and oe-NC group). Moreover, TE-1 cells treated with restored HOTAIR were added with either 5-Aza-CdR or DMSO. ( a ) MTHFR mRNA level in EC tissues ( n = 70) and adjacent normal tissues (n = 70) determined by RT-qPCR, normalized to GAPDH. ( b ) Distribution of CpG islands within MTHFR promoter region. ( c ) MSP was adopted to detect methylation level of MTHFR promoter region in EC tissues (n = 70) and adjacent normal tissues (n = 70). ( d ) The binding sites between the lncRNA HOTAIR and MTHFR promoter regions predicted using Blast comparison website. ( e ) Target relationship verified by <t>dual</t> <t>luciferase</t> <t>reporter</t> gene <t>assay.</t> ( f ) RIP was utilized to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) affected by HOTAIR in TE-1 cells. ( g ) CHIP was employed to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) within MTHFR promoter region in TE-1 cells. ( h ), MSP was used to detect methylation level of TE-1 cells. ( i ) MTHFR mRNA level in TE-1 cells after restoration or depletion of HOTAIR determined by RT-qPCR, normalized to GAPDH. MTHFR (75kD), GAPDH (36kD) ( j ) MTHFR protein level in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. ( k ) Protein band patterns of MTHFR in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by paired t test between EC tissues and adjacent normal tissues, by unpaired t test between EC cells and HEEC cells, and by ANOVA followed by Tukey’s post hoc test among three or more groups. * p
    Dual Luciferase Reporter Assay System, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 92/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SABiosciences dual luciferase reporter assay system
    HOTAIR modulates DNA methylation within MTHFR promoter region. shRNA was delivered into TE-1 cells to specifically blunt HOTAIR (sh-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1 cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells were served as negative controls (sh-NC group and oe-NC group). Moreover, TE-1 cells treated with restored HOTAIR were added with either 5-Aza-CdR or DMSO. ( a ) MTHFR mRNA level in EC tissues ( n = 70) and adjacent normal tissues (n = 70) determined by RT-qPCR, normalized to GAPDH. ( b ) Distribution of CpG islands within MTHFR promoter region. ( c ) MSP was adopted to detect methylation level of MTHFR promoter region in EC tissues (n = 70) and adjacent normal tissues (n = 70). ( d ) The binding sites between the lncRNA HOTAIR and MTHFR promoter regions predicted using Blast comparison website. ( e ) Target relationship verified by <t>dual</t> <t>luciferase</t> <t>reporter</t> gene <t>assay.</t> ( f ) RIP was utilized to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) affected by HOTAIR in TE-1 cells. ( g ) CHIP was employed to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) within MTHFR promoter region in TE-1 cells. ( h ), MSP was used to detect methylation level of TE-1 cells. ( i ) MTHFR mRNA level in TE-1 cells after restoration or depletion of HOTAIR determined by RT-qPCR, normalized to GAPDH. MTHFR (75kD), GAPDH (36kD) ( j ) MTHFR protein level in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. ( k ) Protein band patterns of MTHFR in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by paired t test between EC tissues and adjacent normal tissues, by unpaired t test between EC cells and HEEC cells, and by ANOVA followed by Tukey’s post hoc test among three or more groups. * p
    Dual Luciferase Reporter Assay System, supplied by SABiosciences, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MEGA Inc dual luciferase reporter assay system
    HOTAIR modulates DNA methylation within MTHFR promoter region. shRNA was delivered into TE-1 cells to specifically blunt HOTAIR (sh-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1 cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells were served as negative controls (sh-NC group and oe-NC group). Moreover, TE-1 cells treated with restored HOTAIR were added with either 5-Aza-CdR or DMSO. ( a ) MTHFR mRNA level in EC tissues ( n = 70) and adjacent normal tissues (n = 70) determined by RT-qPCR, normalized to GAPDH. ( b ) Distribution of CpG islands within MTHFR promoter region. ( c ) MSP was adopted to detect methylation level of MTHFR promoter region in EC tissues (n = 70) and adjacent normal tissues (n = 70). ( d ) The binding sites between the lncRNA HOTAIR and MTHFR promoter regions predicted using Blast comparison website. ( e ) Target relationship verified by <t>dual</t> <t>luciferase</t> <t>reporter</t> gene <t>assay.</t> ( f ) RIP was utilized to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) affected by HOTAIR in TE-1 cells. ( g ) CHIP was employed to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) within MTHFR promoter region in TE-1 cells. ( h ), MSP was used to detect methylation level of TE-1 cells. ( i ) MTHFR mRNA level in TE-1 cells after restoration or depletion of HOTAIR determined by RT-qPCR, normalized to GAPDH. MTHFR (75kD), GAPDH (36kD) ( j ) MTHFR protein level in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. ( k ) Protein band patterns of MTHFR in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by paired t test between EC tissues and adjacent normal tissues, by unpaired t test between EC cells and HEEC cells, and by ANOVA followed by Tukey’s post hoc test among three or more groups. * p
    Dual Luciferase Reporter Assay System, supplied by MEGA Inc, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    TaKaRa dual luciferase reporter assay system
    MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b <t>Dual</t> <t>luciferase</t> <t>reporter</t> <t>assay</t> confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p
    Dual Luciferase Reporter Assay System, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cayman Chemical dual luciferase reporter assay system
    MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b <t>Dual</t> <t>luciferase</t> <t>reporter</t> <t>assay</t> confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p
    Dual Luciferase Reporter Assay System, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co dual luciferase reporter assay system
    MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b <t>Dual</t> <t>luciferase</t> <t>reporter</t> <t>assay</t> confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p
    Dual Luciferase Reporter Assay System, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fisher Scientific luciferase assay dual luciferase reporter system psichecktm
    MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b <t>Dual</t> <t>luciferase</t> <t>reporter</t> <t>assay</t> confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p
    Luciferase Assay Dual Luciferase Reporter System Psichecktm, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Promega dual luciferase reporter assays system
    YAP positively regulates Slug transcription. A. Western blot <t>assays</t> show YAP protein expression in different transfection conditions, and qPCR to detect Slug mRNA expression under different conditions of YAP expression. B. <t>Dual-luciferase</t> assay was used to detect up-regulation level of Slug transcriptional activity in stable SW620 cell line overexpressing YAP. Dual-luciferase <t>reporter</t> assays confirm that YAP can positively active the transcription of Slug-luc. *P
    Dual Luciferase Reporter Assays System, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega luciferase reporter assay a dual luciferase reporter assay system
    YAP positively regulates Slug transcription. A. Western blot <t>assays</t> show YAP protein expression in different transfection conditions, and qPCR to detect Slug mRNA expression under different conditions of YAP expression. B. <t>Dual-luciferase</t> assay was used to detect up-regulation level of Slug transcriptional activity in stable SW620 cell line overexpressing YAP. Dual-luciferase <t>reporter</t> assays confirm that YAP can positively active the transcription of Slug-luc. *P
    Luciferase Reporter Assay A Dual Luciferase Reporter Assay System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PRL-3 transcriptionally regulates the expression of RAP1. Notes: ( A ) Western blot results showed that overexpression of PRL-3 increased the RAP1 level (left), and silencing of PRL-3 decreased RAP1 level (right). ( B ) RT-PCR results showed that overexpression of PRL-3 increased RAP1 level (upper) and silencing of PRL-3 decreased RAP1 level (lower). ( C ) Dual luciferase reporter assays of rap1 gene promoter in HCT116 and 293 T cells expressing pcDNA3.0-myc-NC/PRL-3 and pGLB-RAP1(−1,500/+150)/(−500/+150). Results were normalized to the activity obtained in cells transfected with pcDNA3.0-myc-NC and pGLB-RAP1 (−1,500/+150). ( D ) ChIP assay using PRL-3 or IgG in HCT116 cells, and qPCR of different promoter regions of rap1 gene. Values correspond to the ratio between the PRL-3 antibody immunoprecipitated DNA with respect to the IgG immunoprecipitated DNA. ( E ) Gene expression correlation analysis between PRL-3 and RAP1 in TCGA data ( P =0.0005, R =0.52) and GTEx data ( P

    Journal: Cancer Management and Research

    Article Title: Knockdown of PRL-3 increases mitochondrial superoxide anion production through transcriptional regulation of RAP1

    doi: 10.2147/CMAR.S165344

    Figure Lengend Snippet: PRL-3 transcriptionally regulates the expression of RAP1. Notes: ( A ) Western blot results showed that overexpression of PRL-3 increased the RAP1 level (left), and silencing of PRL-3 decreased RAP1 level (right). ( B ) RT-PCR results showed that overexpression of PRL-3 increased RAP1 level (upper) and silencing of PRL-3 decreased RAP1 level (lower). ( C ) Dual luciferase reporter assays of rap1 gene promoter in HCT116 and 293 T cells expressing pcDNA3.0-myc-NC/PRL-3 and pGLB-RAP1(−1,500/+150)/(−500/+150). Results were normalized to the activity obtained in cells transfected with pcDNA3.0-myc-NC and pGLB-RAP1 (−1,500/+150). ( D ) ChIP assay using PRL-3 or IgG in HCT116 cells, and qPCR of different promoter regions of rap1 gene. Values correspond to the ratio between the PRL-3 antibody immunoprecipitated DNA with respect to the IgG immunoprecipitated DNA. ( E ) Gene expression correlation analysis between PRL-3 and RAP1 in TCGA data ( P =0.0005, R =0.52) and GTEx data ( P

    Article Snippet: Luciferase activity was measured using the Dual-Luciferase® Reporter Assay System (Thermo Fisher Scientific).

    Techniques: Expressing, Western Blot, Over Expression, Reverse Transcription Polymerase Chain Reaction, Luciferase, Activity Assay, Transfection, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation

    PDGFRB is a direct target gene of miR-518b. (A) Inverse relationship between the expression of miR-518b and PDGFRB was detected in GBM specimens (magnification, 200x). (B) Western blot analysis of PDGFRB expression in U87 and U251 cells 48 h post-transfection with miR-518b and miR-ctrl. (C) PDGFRB mRNA 3′UTR contains binding sites for miR-518b. (D) U87 and U251 cells were co-transfected with the dual-luciferase reporter plasmid carrying the Wt or Mut 3′UTR sequences of PDGFRB and miR-518b or miR-ctrl mimics. A luciferase reporter system analysis was performed. Data are presented as the mean ± standard deviation (n=5). *P

    Journal: Molecular Medicine Reports

    Article Title: MicroRNA-518b functions as a tumor suppressor in glioblastoma by targeting PDGFRB

    doi: 10.3892/mmr.2017.7298

    Figure Lengend Snippet: PDGFRB is a direct target gene of miR-518b. (A) Inverse relationship between the expression of miR-518b and PDGFRB was detected in GBM specimens (magnification, 200x). (B) Western blot analysis of PDGFRB expression in U87 and U251 cells 48 h post-transfection with miR-518b and miR-ctrl. (C) PDGFRB mRNA 3′UTR contains binding sites for miR-518b. (D) U87 and U251 cells were co-transfected with the dual-luciferase reporter plasmid carrying the Wt or Mut 3′UTR sequences of PDGFRB and miR-518b or miR-ctrl mimics. A luciferase reporter system analysis was performed. Data are presented as the mean ± standard deviation (n=5). *P

    Article Snippet: After 24 h, luciferase activity was measured using the Dual-Luciferase Reporter Assay system (Beyotime Institute of Biotechnology, Beijing, China) according to the manufacturer's protocol.

    Techniques: Expressing, Western Blot, Transfection, Binding Assay, Luciferase, Plasmid Preparation, Standard Deviation

    miR-155 directly targeted on MafB. (a) The target site of miR-155 and MafB 3′UTR was measured based on TargetScan and microRNA.org. (b) Investigation of the regulatory effect of miR-155 on MafB expression using dual-luciferase reporter assay in 293T cells. (c) and (d and e) The expression of MafB in OGD/R-induced SH-SY5Y cells transfected with miR-155 or anti-miR-155 was measured through qRT-PCR and western blot analyses respectively ∗ P

    Journal: BioMed Research International

    Article Title: miR-155 Knockdown Protects against Cerebral Ischemia and Reperfusion Injury by Targeting MafB

    doi: 10.1155/2020/6458204

    Figure Lengend Snippet: miR-155 directly targeted on MafB. (a) The target site of miR-155 and MafB 3′UTR was measured based on TargetScan and microRNA.org. (b) Investigation of the regulatory effect of miR-155 on MafB expression using dual-luciferase reporter assay in 293T cells. (c) and (d and e) The expression of MafB in OGD/R-induced SH-SY5Y cells transfected with miR-155 or anti-miR-155 was measured through qRT-PCR and western blot analyses respectively ∗ P

    Article Snippet: After culturing for 48 h, we collected the cells and detected the luciferase activity with a Dual-Luciferase® Reporter Assay System (TransGen Biotech, Beijing, China) according to the standard protocols and the associate luciferase activity was evaluated as the ratio of firefly luminescence to Renilla luminescence.

    Techniques: Expressing, Luciferase, Reporter Assay, Transfection, Quantitative RT-PCR, Western Blot

    HOTAIR modulates DNA methylation within MTHFR promoter region. shRNA was delivered into TE-1 cells to specifically blunt HOTAIR (sh-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1 cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells were served as negative controls (sh-NC group and oe-NC group). Moreover, TE-1 cells treated with restored HOTAIR were added with either 5-Aza-CdR or DMSO. ( a ) MTHFR mRNA level in EC tissues ( n = 70) and adjacent normal tissues (n = 70) determined by RT-qPCR, normalized to GAPDH. ( b ) Distribution of CpG islands within MTHFR promoter region. ( c ) MSP was adopted to detect methylation level of MTHFR promoter region in EC tissues (n = 70) and adjacent normal tissues (n = 70). ( d ) The binding sites between the lncRNA HOTAIR and MTHFR promoter regions predicted using Blast comparison website. ( e ) Target relationship verified by dual luciferase reporter gene assay. ( f ) RIP was utilized to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) affected by HOTAIR in TE-1 cells. ( g ) CHIP was employed to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) within MTHFR promoter region in TE-1 cells. ( h ), MSP was used to detect methylation level of TE-1 cells. ( i ) MTHFR mRNA level in TE-1 cells after restoration or depletion of HOTAIR determined by RT-qPCR, normalized to GAPDH. MTHFR (75kD), GAPDH (36kD) ( j ) MTHFR protein level in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. ( k ) Protein band patterns of MTHFR in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by paired t test between EC tissues and adjacent normal tissues, by unpaired t test between EC cells and HEEC cells, and by ANOVA followed by Tukey’s post hoc test among three or more groups. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: LncRNA HOTAIR-mediated MTHFR methylation inhibits 5-fluorouracil sensitivity in esophageal cancer cells

    doi: 10.1186/s13046-020-01610-1

    Figure Lengend Snippet: HOTAIR modulates DNA methylation within MTHFR promoter region. shRNA was delivered into TE-1 cells to specifically blunt HOTAIR (sh-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1 cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells were served as negative controls (sh-NC group and oe-NC group). Moreover, TE-1 cells treated with restored HOTAIR were added with either 5-Aza-CdR or DMSO. ( a ) MTHFR mRNA level in EC tissues ( n = 70) and adjacent normal tissues (n = 70) determined by RT-qPCR, normalized to GAPDH. ( b ) Distribution of CpG islands within MTHFR promoter region. ( c ) MSP was adopted to detect methylation level of MTHFR promoter region in EC tissues (n = 70) and adjacent normal tissues (n = 70). ( d ) The binding sites between the lncRNA HOTAIR and MTHFR promoter regions predicted using Blast comparison website. ( e ) Target relationship verified by dual luciferase reporter gene assay. ( f ) RIP was utilized to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) affected by HOTAIR in TE-1 cells. ( g ) CHIP was employed to detect the enrichment of DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) within MTHFR promoter region in TE-1 cells. ( h ), MSP was used to detect methylation level of TE-1 cells. ( i ) MTHFR mRNA level in TE-1 cells after restoration or depletion of HOTAIR determined by RT-qPCR, normalized to GAPDH. MTHFR (75kD), GAPDH (36kD) ( j ) MTHFR protein level in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. ( k ) Protein band patterns of MTHFR in TE-1 cells after restoration or depletion of HOTAIR determined by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by paired t test between EC tissues and adjacent normal tissues, by unpaired t test between EC cells and HEEC cells, and by ANOVA followed by Tukey’s post hoc test among three or more groups. * p

    Article Snippet: The Dual-Luciferase Reporter Assay System from Genecopoeia (D0010, Beijing Solarbio Science & Technology Co. Ltd., Beijing, China) was employed to detect the luciferase activity of MTHFR promoter region induced by lncRNA HOTAIR in EC cells.

    Techniques: DNA Methylation Assay, shRNA, Expressing, Quantitative RT-PCR, Methylation, Binding Assay, Luciferase, Reporter Gene Assay, Chromatin Immunoprecipitation, Western Blot

    MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b Dual luciferase reporter assay confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p

    Journal: Molecular Medicine

    Article Title: Long non-coding RNA MCM3AP-AS1 promotes growth and migration through modulating FOXK1 by sponging miR-138-5p in pancreatic cancer

    doi: 10.1186/s10020-019-0121-2

    Figure Lengend Snippet: MCM3AP-AS1 promoted FOXK1 expression by targeting miR-138-5p. a Bioinformatic analysis predicted that miR-138-5p could target FOXK1. b Dual luciferase reporter assay confirmed the relationship between miR-138-5p and FOXK1. c qRT-PCR detected miR-138-5p level. d - f qRT-PCR and western blot analysis measured FOXK1 level. g The correlation between miR-138-5p and FOXK1 was identified. (H) FOXK1 level was examined using quantitative real time PCR and western blot analysis. n = 3. (**, p

    Article Snippet: 48 h post-transfection, dual-luciferase reporter assay system (Takara, Dalian, China) was used to examine relative luciferase activities.

    Techniques: Expressing, Luciferase, Reporter Assay, Quantitative RT-PCR, Western Blot, Real-time Polymerase Chain Reaction

    MCM3AP-AS1 was negatively associated with miR-138-5p. a Bioinformatic analysis predicted that MCM3AP-AS1 could sponge miR-138-5p. b qRT-PCR detected miR-138-5p level. c Dual luciferase reporter assay demonstrated the interaction between MCM3AP-AS1 and miR-138-5p. d The binding of MCM3AP-AS1 and miR-138-5p was identified via RIP assay. e - f qRT-PCR analyzed miR-138-5p level. g The correlation between MCM3AP-AS1 and miR-138-5p was assayed. n = 3. (**, p

    Journal: Molecular Medicine

    Article Title: Long non-coding RNA MCM3AP-AS1 promotes growth and migration through modulating FOXK1 by sponging miR-138-5p in pancreatic cancer

    doi: 10.1186/s10020-019-0121-2

    Figure Lengend Snippet: MCM3AP-AS1 was negatively associated with miR-138-5p. a Bioinformatic analysis predicted that MCM3AP-AS1 could sponge miR-138-5p. b qRT-PCR detected miR-138-5p level. c Dual luciferase reporter assay demonstrated the interaction between MCM3AP-AS1 and miR-138-5p. d The binding of MCM3AP-AS1 and miR-138-5p was identified via RIP assay. e - f qRT-PCR analyzed miR-138-5p level. g The correlation between MCM3AP-AS1 and miR-138-5p was assayed. n = 3. (**, p

    Article Snippet: 48 h post-transfection, dual-luciferase reporter assay system (Takara, Dalian, China) was used to examine relative luciferase activities.

    Techniques: Quantitative RT-PCR, Luciferase, Reporter Assay, Binding Assay

    YAP positively regulates Slug transcription. A. Western blot assays show YAP protein expression in different transfection conditions, and qPCR to detect Slug mRNA expression under different conditions of YAP expression. B. Dual-luciferase assay was used to detect up-regulation level of Slug transcriptional activity in stable SW620 cell line overexpressing YAP. Dual-luciferase reporter assays confirm that YAP can positively active the transcription of Slug-luc. *P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: YAP promotes epithelial mesenchymal transition by upregulating Slug expression in human colorectal cancer cells

    doi:

    Figure Lengend Snippet: YAP positively regulates Slug transcription. A. Western blot assays show YAP protein expression in different transfection conditions, and qPCR to detect Slug mRNA expression under different conditions of YAP expression. B. Dual-luciferase assay was used to detect up-regulation level of Slug transcriptional activity in stable SW620 cell line overexpressing YAP. Dual-luciferase reporter assays confirm that YAP can positively active the transcription of Slug-luc. *P

    Article Snippet: After 40 h, cell extracts were prepared and reporter activity was determined using the dual-luciferase reporter assays system purchased from Promega (E1910, USA).

    Techniques: Western Blot, Expressing, Transfection, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay