dtt Promega Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore dtt
    Dtt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 36382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt/product/Millipore
    Average 99 stars, based on 36382 article reviews
    Price from $9.99 to $1999.99
    dtt - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    94
    Promega dithiothreitol dtt
    Dithiothreitol Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 1939 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dithiothreitol dtt/product/Promega
    Average 94 stars, based on 1939 article reviews
    Price from $9.99 to $1999.99
    dithiothreitol dtt - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    94
    Promega molecular grade dithiothreitol dtt
    Drug-induced ER stress upregulates DmManf expression. A–B) In Schneider 2 (S2) cells, ER stress was induced by thapsigargin (TG), tunicamycin (TM) and dithiothreitol <t>(DTT).</t> <t>DMSO</t> was used as a control treatment. A) The mRNA levels of DmManf and Hsc3 were analysed by qPCR, values were normalised to control treatment (DMSO). B) RT-PCR and agarose gel electrophoresis analysis revealed two transcripts of Xbp1 , unspliced ( Xbp1 u ) and spliced ( Xbp1 s ). RpL32 was used as a loading control. C–D) qPCR analysis of Hsc3 and Xbp1 expression in DmManf mutant (C) and DmManf overexpressing (D) larvae. Expression of Hsc3 was not altered but Xbp1s mRNA level was increased in response to overexpression of DmManf . The overexpression of DmManf resulted in 165-fold increase in DmManf mRNA level (±23, P
    Molecular Grade Dithiothreitol Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/molecular grade dithiothreitol dtt/product/Promega
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    molecular grade dithiothreitol dtt - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    93
    Promega dithiothreitol
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dithiothreitol, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 5529 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dithiothreitol/product/Promega
    Average 93 stars, based on 5529 article reviews
    Price from $9.99 to $1999.99
    dithiothreitol - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    88
    Promega 1 4 dithiothreotol dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    1 4 Dithiothreotol Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1 4 dithiothreotol dtt/product/Promega
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    1 4 dithiothreotol dtt - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    85
    Promega dtt promega solution
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dtt Promega Solution, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt promega solution/product/Promega
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    dtt promega solution - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    88
    Promega dl dithiothretiol dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dl Dithiothretiol Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dl dithiothretiol dtt/product/Promega
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dl dithiothretiol dtt - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    80
    Promega rnasin dithiothreitol dtt water
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Rnasin Dithiothreitol Dtt Water, supplied by Promega, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnasin dithiothreitol dtt water/product/Promega
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rnasin dithiothreitol dtt water - by Bioz Stars, 2020-07
    80/100 stars
      Buy from Supplier

    92
    Promega m dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    M Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 628 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m dtt/product/Promega
    Average 92 stars, based on 628 article reviews
    Price from $9.99 to $1999.99
    m dtt - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    89
    Promega 1m dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    1m Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1m dtt/product/Promega
    Average 89 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    1m dtt - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    91
    Promega dtt free promega passive lysis buffer
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dtt Free Promega Passive Lysis Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt free promega passive lysis buffer/product/Promega
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dtt free promega passive lysis buffer - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    94
    New England Biolabs dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dtt, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 4112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt/product/New England Biolabs
    Average 94 stars, based on 4112 article reviews
    Price from $9.99 to $1999.99
    dtt - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    85
    Promega dtt iaa
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dtt Iaa, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt iaa/product/Promega
    Average 85 stars, based on 27 article reviews
    Price from $9.99 to $1999.99
    dtt iaa - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    85
    Promega mmol dtt
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Mmol Dtt, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmol dtt/product/Promega
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    mmol dtt - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    89
    Promega dtt buffer
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    Dtt Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt buffer/product/Promega
    Average 89 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    dtt buffer - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    92
    Promega 1m dithiothreitol
    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM <t>dithiothreitol-20</t> U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.
    1m Dithiothreitol, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1m dithiothreitol/product/Promega
    Average 92 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    1m dithiothreitol - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    Image Search Results


    Drug-induced ER stress upregulates DmManf expression. A–B) In Schneider 2 (S2) cells, ER stress was induced by thapsigargin (TG), tunicamycin (TM) and dithiothreitol (DTT). DMSO was used as a control treatment. A) The mRNA levels of DmManf and Hsc3 were analysed by qPCR, values were normalised to control treatment (DMSO). B) RT-PCR and agarose gel electrophoresis analysis revealed two transcripts of Xbp1 , unspliced ( Xbp1 u ) and spliced ( Xbp1 s ). RpL32 was used as a loading control. C–D) qPCR analysis of Hsc3 and Xbp1 expression in DmManf mutant (C) and DmManf overexpressing (D) larvae. Expression of Hsc3 was not altered but Xbp1s mRNA level was increased in response to overexpression of DmManf . The overexpression of DmManf resulted in 165-fold increase in DmManf mRNA level (±23, P

    Journal: PLoS ONE

    Article Title: Exploring the Conserved Role of MANF in the Unfolded Protein Response in Drosophila melanogaster

    doi: 10.1371/journal.pone.0151550

    Figure Lengend Snippet: Drug-induced ER stress upregulates DmManf expression. A–B) In Schneider 2 (S2) cells, ER stress was induced by thapsigargin (TG), tunicamycin (TM) and dithiothreitol (DTT). DMSO was used as a control treatment. A) The mRNA levels of DmManf and Hsc3 were analysed by qPCR, values were normalised to control treatment (DMSO). B) RT-PCR and agarose gel electrophoresis analysis revealed two transcripts of Xbp1 , unspliced ( Xbp1 u ) and spliced ( Xbp1 s ). RpL32 was used as a loading control. C–D) qPCR analysis of Hsc3 and Xbp1 expression in DmManf mutant (C) and DmManf overexpressing (D) larvae. Expression of Hsc3 was not altered but Xbp1s mRNA level was increased in response to overexpression of DmManf . The overexpression of DmManf resulted in 165-fold increase in DmManf mRNA level (±23, P

    Article Snippet: Cells were treated with DMSO, 1 μM thapsigargin (Molecular Probes), 1 mM DTT (Promega) or 10 μg/ml tunicamycin for 20 hours, collected and total RNA was extracted with NucleoSpin® RNA II (Macherey-Nagel).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Mutagenesis, Over Expression

    G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM dithiothreitol-20 U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.

    Journal: Journal of Virology

    Article Title: Selective Translation of Eukaryotic mRNAs: Functional Molecular Analysis of GRSF-1, a Positive Regulator of Influenza Virus Protein Synthesis

    doi: 10.1128/JVI.76.20.10417-10426.2002

    Figure Lengend Snippet: G13 and U14 in the NS1 5′ UTR are required for GRSF-1 binding. Wild-type (Wt) GST-GRSF-1 (1.25 × 10 −11 mol) was incubated with 10 6 dpm of radiolabeled wild-type NS1 5′ UTR RNA in the absence or presence of increasing concentrations (10- or 100-fold molar excess) of unlabeled wild-type (lanes 2 to 4), U14C (lanes 5 to 7), or G13C/U14C (lanes 8 to 10) competitor RNAs in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM dithiothreitol-20 U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of three independent experiments.

    Article Snippet: The S10 lysate was supplemented with 0.0156 mg of tRNA per ml, 0.62 mM ATP, 0.037 mM GTP, 6.22 mM creatine phosphate, 0.0156 mg of creatine kinase per ml, 11.8 mM HEPES (pH 7.6), 1.24 mM dithiothreitol, 15.6 μM complete amino acid mixture (Promega), and 0.156 mM spermidine.

    Techniques: Binding Assay, Incubation, Polyacrylamide Gel Electrophoresis

    GRSF-1 interacts with specific sequences within the influenza virus mRNA 5′ UTR as detected by gel mobility shift analysis. Recombinant GRSF-1 (100 ng) was incubated with radiolabeled NP-c, NPD3, NPD6, NPD9, NP12, NP14, M1, NA, PB1, and NP-B RNAs (10 5 dpm) in buffer that contained 5 mM HEPES (pH 7.6), 25 mM KCl, 2 mM MgCl 2 , 3.8% glycerol, 100 mM NaCl, 0.02 mM dithiothreitol, 2 mM GTP, and 1.5 mM ATP at 30°C for 20 min. The resulting RNA-protein complexes were resolved by 5% native PAGE and visualized by autoradiography. These data are representative of two independent experiments.

    Journal: Journal of Virology

    Article Title: Selective Translation of Eukaryotic mRNAs: Functional Molecular Analysis of GRSF-1, a Positive Regulator of Influenza Virus Protein Synthesis

    doi: 10.1128/JVI.76.20.10417-10426.2002

    Figure Lengend Snippet: GRSF-1 interacts with specific sequences within the influenza virus mRNA 5′ UTR as detected by gel mobility shift analysis. Recombinant GRSF-1 (100 ng) was incubated with radiolabeled NP-c, NPD3, NPD6, NPD9, NP12, NP14, M1, NA, PB1, and NP-B RNAs (10 5 dpm) in buffer that contained 5 mM HEPES (pH 7.6), 25 mM KCl, 2 mM MgCl 2 , 3.8% glycerol, 100 mM NaCl, 0.02 mM dithiothreitol, 2 mM GTP, and 1.5 mM ATP at 30°C for 20 min. The resulting RNA-protein complexes were resolved by 5% native PAGE and visualized by autoradiography. These data are representative of two independent experiments.

    Article Snippet: The S10 lysate was supplemented with 0.0156 mg of tRNA per ml, 0.62 mM ATP, 0.037 mM GTP, 6.22 mM creatine phosphate, 0.0156 mg of creatine kinase per ml, 11.8 mM HEPES (pH 7.6), 1.24 mM dithiothreitol, 15.6 μM complete amino acid mixture (Promega), and 0.156 mM spermidine.

    Techniques: Mobility Shift, Recombinant, Incubation, Clear Native PAGE, Autoradiography

    Second RRM of GRSF-1 is required for mRNA binding in vitro. (A) Coomassie blue-stained denaturing 12% polyacrylamide gel of equal moles (1.25 × 10 −11 mol) of wild-type and mutant GST-GRSF-1 fusion proteins purified by glutathione affinity chromatography from E. coli . (B) UV cross-linking analysis of thrombin-cleaved GRSF-1 (rGRSF-1) and intact wild-type (Wt) and mutant GST-GRSF-1 fusion proteins (1.25 × 10 −11 mol) incubated with radiolabeled NP 5′ UTR RNA (10 6 dpm) in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM dithiothreitol-20 U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of two independent experiments.

    Journal: Journal of Virology

    Article Title: Selective Translation of Eukaryotic mRNAs: Functional Molecular Analysis of GRSF-1, a Positive Regulator of Influenza Virus Protein Synthesis

    doi: 10.1128/JVI.76.20.10417-10426.2002

    Figure Lengend Snippet: Second RRM of GRSF-1 is required for mRNA binding in vitro. (A) Coomassie blue-stained denaturing 12% polyacrylamide gel of equal moles (1.25 × 10 −11 mol) of wild-type and mutant GST-GRSF-1 fusion proteins purified by glutathione affinity chromatography from E. coli . (B) UV cross-linking analysis of thrombin-cleaved GRSF-1 (rGRSF-1) and intact wild-type (Wt) and mutant GST-GRSF-1 fusion proteins (1.25 × 10 −11 mol) incubated with radiolabeled NP 5′ UTR RNA (10 6 dpm) in 5 mM HEPES (pH 7.6)-25 mM KCl-2 mM MgCl 2 -5% glycerol-100 mM NaCl-2 mM dithiothreitol-20 U of RNasin (Promega) at 30°C for 15 min. Samples were then exposed to UV light (1.0 J) in a Stratalinker (Stratagene), followed by incubation with 10 mg of RNase A per ml and 2 U of RNase T 1 at 37°C for 30 min. Samples were suspended in 1× Laemmli buffer and separated by SDS-10% PAGE, and the gel was dried and visualized with a Storm 850 phosphorimager (Molecular Dynamics). These data are representative of two independent experiments.

    Article Snippet: The S10 lysate was supplemented with 0.0156 mg of tRNA per ml, 0.62 mM ATP, 0.037 mM GTP, 6.22 mM creatine phosphate, 0.0156 mg of creatine kinase per ml, 11.8 mM HEPES (pH 7.6), 1.24 mM dithiothreitol, 15.6 μM complete amino acid mixture (Promega), and 0.156 mM spermidine.

    Techniques: Binding Assay, In Vitro, Staining, Mutagenesis, Purification, Affinity Chromatography, Incubation, Polyacrylamide Gel Electrophoresis

    GRSF-1 binds the conserved A-box of the influenza virus NP 5′ UTR. Shown here is a representative autoradiogram of RNA gel shift analysis of radiolabeled influenza virus NP (lanes 1 to 7) or NP-A (lanes 8 to 14) 5′ UTR RNA incubated with increasing concentrations (50, 100, and 200 ng) of thrombin-cleaved (rGRSF-1) or uncleaved GST-GRSF-1. Proteins were incubated with radiolabeled RNA (100,000 dpm) in buffer that contained 5 mM HEPES (pH 7.6), 25 mM KCl, 2 mM MgCl 2 , 3.8% glycerol, 100 mM NaCl, 0.02 mM dithiothreitol, 2 mM GTP, and 1.5 mM ATP at 30°C for 20 min. Samples were then electrophoresed on a 5% polyacrylamide gel at 4°C and visualized by autoradiography. These data are representative of two independent experiments.

    Journal: Journal of Virology

    Article Title: Selective Translation of Eukaryotic mRNAs: Functional Molecular Analysis of GRSF-1, a Positive Regulator of Influenza Virus Protein Synthesis

    doi: 10.1128/JVI.76.20.10417-10426.2002

    Figure Lengend Snippet: GRSF-1 binds the conserved A-box of the influenza virus NP 5′ UTR. Shown here is a representative autoradiogram of RNA gel shift analysis of radiolabeled influenza virus NP (lanes 1 to 7) or NP-A (lanes 8 to 14) 5′ UTR RNA incubated with increasing concentrations (50, 100, and 200 ng) of thrombin-cleaved (rGRSF-1) or uncleaved GST-GRSF-1. Proteins were incubated with radiolabeled RNA (100,000 dpm) in buffer that contained 5 mM HEPES (pH 7.6), 25 mM KCl, 2 mM MgCl 2 , 3.8% glycerol, 100 mM NaCl, 0.02 mM dithiothreitol, 2 mM GTP, and 1.5 mM ATP at 30°C for 20 min. Samples were then electrophoresed on a 5% polyacrylamide gel at 4°C and visualized by autoradiography. These data are representative of two independent experiments.

    Article Snippet: The S10 lysate was supplemented with 0.0156 mg of tRNA per ml, 0.62 mM ATP, 0.037 mM GTP, 6.22 mM creatine phosphate, 0.0156 mg of creatine kinase per ml, 11.8 mM HEPES (pH 7.6), 1.24 mM dithiothreitol, 15.6 μM complete amino acid mixture (Promega), and 0.156 mM spermidine.

    Techniques: Electrophoretic Mobility Shift Assay, Incubation, Autoradiography