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    Jena Bioscience utp
    Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and <t>UTP</t> at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 <t>P-CTP</t> (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p

    https://www.bioz.com/result/utp/product/Jena Bioscience
    Average 95 stars, based on 1 article reviews
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    utp - by Bioz Stars, 2021-06
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    1) Product Images from "Guanosine inhibits hepatitis C virus replication and increases indel frequencies, associated with altered intracellular nucleotide pools"

    Article Title: Guanosine inhibits hepatitis C virus replication and increases indel frequencies, associated with altered intracellular nucleotide pools

    Journal: bioRxiv

    doi: 10.1101/2020.02.21.959536

    Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and UTP at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 P-CTP (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p
    Figure Legend Snippet: Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and UTP at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 P-CTP (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p

    Techniques Used: Activity Assay, Concentration Assay

    Effect of Gua on NS5BΔ21 RNA polymerase activity. (A) Recombinant HCV NS5B Δ 21 polymerase was added to a reaction containing a 540-nt RNA template [ 18 ], the four nucleoside-triphosphates (ATP, CTP, GTP, and UTP) and the indicated concentrations of Gua. Product quantification from three replicates (average ± SEM) and a representative experiment (below) are shown. Polymerase activity is normalized with respect to its maximum activity. The band indicates a new synthesis RNA product of 540 nt length. (B) A representative experiment as in A, but using the 19-nt LE19 RNA as a template. DN, PE, and TS indicate reaction products of de novo synthesis, primer extension, and template switching, respectively [ 54 ]. Procedures are detailed in Materials and Methods. Significance (Student’s T-test): ** p
    Figure Legend Snippet: Effect of Gua on NS5BΔ21 RNA polymerase activity. (A) Recombinant HCV NS5B Δ 21 polymerase was added to a reaction containing a 540-nt RNA template [ 18 ], the four nucleoside-triphosphates (ATP, CTP, GTP, and UTP) and the indicated concentrations of Gua. Product quantification from three replicates (average ± SEM) and a representative experiment (below) are shown. Polymerase activity is normalized with respect to its maximum activity. The band indicates a new synthesis RNA product of 540 nt length. (B) A representative experiment as in A, but using the 19-nt LE19 RNA as a template. DN, PE, and TS indicate reaction products of de novo synthesis, primer extension, and template switching, respectively [ 54 ]. Procedures are detailed in Materials and Methods. Significance (Student’s T-test): ** p

    Techniques Used: Activity Assay, Recombinant

    Effect of nucleoside-triphosphate concentration on NS5BΔ21 RNA polymerase activity. ( A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (100, 500, 800, and 1000 μM) of UTP in the presence of radiolabeled α 32 P-CTP. ATP and GTP concentrations were maintained at 100 μM and 500 μM, respectively (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ATP, with UTP and GTP maintained at 100 μM and 500 μM, respectively. (C) Corresponds to experiments as in A but for increasing concentrations of GTP, with ATP and UTP both maintained at 100 μM. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between the activity values that link, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): *** p
    Figure Legend Snippet: Effect of nucleoside-triphosphate concentration on NS5BΔ21 RNA polymerase activity. ( A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (100, 500, 800, and 1000 μM) of UTP in the presence of radiolabeled α 32 P-CTP. ATP and GTP concentrations were maintained at 100 μM and 500 μM, respectively (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ATP, with UTP and GTP maintained at 100 μM and 500 μM, respectively. (C) Corresponds to experiments as in A but for increasing concentrations of GTP, with ATP and UTP both maintained at 100 μM. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between the activity values that link, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): *** p

    Techniques Used: Concentration Assay, Activity Assay

    Related Articles

    Enzymatic Assay:

    Article Title: Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni.
    Article Snippet: .. The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. ..

    Concentration Assay:

    Article Title: Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Campylobacter jejuni.
    Article Snippet: .. The enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) catalyses the hydrolysis of dUTP to dUMP and PPi thus controlling the incorporation of uracil into DNA genomes. ..

    Labeling:

    Article Title: Native molecule sequencing by nano-ID reveals synthesis and stability of RNA isoforms
    Article Snippet: IVT of synthetic control RNAs was performed following the manufacturer's instruction. .. For IVT of labeled synthetic RNAs, 100% of UTP (resp. GTP) was substituted with either 5-ethynyl-UTP (5E U; Jena Bioscience), 5-bromo-UTP (5Br U; Sigma-Aldrich), 5-iodo-UTP (5I U; TriLink BioTechnologies), 4-thio-UTP (4S U; Jena Bioscience), or 6-thio-GTP (6S G; Sigma-Aldrich). .. Note that, for performing a successful IVT with 4-thio-UTP and 6-thio-GTP, only a reduction to 80% substitution gave a successful yield.

    Chromatography:

    Article Title: Extinction of Hepatitis C Virus by Ribavirin in Hepatoma Cells Involves Lethal Mutagenesis
    Article Snippet: The separation program started with 22.5 min of an isocratic period with buffer A followed by a linear gradient of 112.5 min to the high concentration buffer 250 mM NH4 H2 PO4 , 500 mM KCl, pH 4.5 (buffer B) and a final isocratic period of 37.5 min with buffer B. .. Prior to sample analysis, 50 µl of 20 pmol/µl UTP, CTP, ATP and GTP, and 80 pmol/µl Ribavirin (Jena Bioscience), were separated to create a processing method using the Waters Empower™ Chromatography Data Software. ..

    Software:

    Article Title: Extinction of Hepatitis C Virus by Ribavirin in Hepatoma Cells Involves Lethal Mutagenesis
    Article Snippet: The separation program started with 22.5 min of an isocratic period with buffer A followed by a linear gradient of 112.5 min to the high concentration buffer 250 mM NH4 H2 PO4 , 500 mM KCl, pH 4.5 (buffer B) and a final isocratic period of 37.5 min with buffer B. .. Prior to sample analysis, 50 µl of 20 pmol/µl UTP, CTP, ATP and GTP, and 80 pmol/µl Ribavirin (Jena Bioscience), were separated to create a processing method using the Waters Empower™ Chromatography Data Software. ..

    Incubation:

    Article Title: Inhibition of dengue virus replication by novel inhibitors of RNA-dependent RNA polymerase and protease activities
    Article Snippet: Briefly, the template was amplified from the cDNA of DENV-2 minus strand 3′-UTR and its RNA was synthesised by the T7 Megascript kit . .. The 100 nM DENV NS5 protein was incubated with 1, 5 or 10 µM test compound, 100 nM (−) 3′-UTR RNA, 20 µM CTP, GTP, UTP and 20 µM BBT-ATP (Jena Bioscience) in a total volume of 30 µL in assay buffer (50 mM Tris-HCl, pH 7.5, 10 mM KCl, 1 mM MgCl2 , 0.3 mM MnCl2 , 0.001% Triton X-100 and 10 µM cysteine) at 25 °C for 60 min. .. The 30 µL of 2.5× stop buffer (200 mM NaCl, 25 mM MgCl2 , 1.5 M diethanolamine, pH 10) with 25 nM calf intestinal alkaline phosphatase (Promega) was added to the reaction after 60-min incubation.

    In Vitro:

    Article Title: The Dynamics of Cytoplasmic mRNA Metabolism
    Article Snippet: Short 5EU standards were prepared by in vitro transcription of annealed DNA oligos to produce a 30 nt and 40 nt RNA, with the latter containing a single 5EU ( ). .. In vitro transcription was performed with the MEGAscript T7 transcription kit (ThermoFisher) according to the manufacturer’s protocol, except UTP was replaced with 5-ethynyluridine-triphosphate (Jena Biosciences) when transcribing the 40 nt RNA. .. Long standards were prepared by in vitro transcription of sequences encoding firefly luciferase and GFP using the MEGAscript T7 transcription kit and 0.1 μM PCR product as the template.

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  • 95
    Jena Bioscience utp
    Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and <t>UTP</t> at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 <t>P-CTP</t> (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p
    Utp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/utp/product/Jena Bioscience
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    utp - by Bioz Stars, 2021-06
    95/100 stars
      Buy from Supplier

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    Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and UTP at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 P-CTP (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p

    Journal: bioRxiv

    Article Title: Guanosine inhibits hepatitis C virus replication and increases indel frequencies, associated with altered intracellular nucleotide pools

    doi: 10.1101/2020.02.21.959536

    Figure Lengend Snippet: Effect of NDPs on NS5BΔ21 RNA polymerase activity. (A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (0, 166, 333, 500, 800, and 1000 μM) of UDP in the presence of ATP and UTP at a final concentration of 100 μM, GTP at 500 μM, and radiolabeled α 32 P-CTP (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ADP. (C) corresponds to experiments as in A but for increasing concentrations of GDP. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between activity values, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): ** p

    Article Snippet: To this end, 50 µl of 20 pmol/µl UTP, CTP, ATP and GTP (Jena Bioscience), were separated prior to sample analysis.

    Techniques: Activity Assay, Concentration Assay

    Effect of Gua on NS5BΔ21 RNA polymerase activity. (A) Recombinant HCV NS5B Δ 21 polymerase was added to a reaction containing a 540-nt RNA template [ 18 ], the four nucleoside-triphosphates (ATP, CTP, GTP, and UTP) and the indicated concentrations of Gua. Product quantification from three replicates (average ± SEM) and a representative experiment (below) are shown. Polymerase activity is normalized with respect to its maximum activity. The band indicates a new synthesis RNA product of 540 nt length. (B) A representative experiment as in A, but using the 19-nt LE19 RNA as a template. DN, PE, and TS indicate reaction products of de novo synthesis, primer extension, and template switching, respectively [ 54 ]. Procedures are detailed in Materials and Methods. Significance (Student’s T-test): ** p

    Journal: bioRxiv

    Article Title: Guanosine inhibits hepatitis C virus replication and increases indel frequencies, associated with altered intracellular nucleotide pools

    doi: 10.1101/2020.02.21.959536

    Figure Lengend Snippet: Effect of Gua on NS5BΔ21 RNA polymerase activity. (A) Recombinant HCV NS5B Δ 21 polymerase was added to a reaction containing a 540-nt RNA template [ 18 ], the four nucleoside-triphosphates (ATP, CTP, GTP, and UTP) and the indicated concentrations of Gua. Product quantification from three replicates (average ± SEM) and a representative experiment (below) are shown. Polymerase activity is normalized with respect to its maximum activity. The band indicates a new synthesis RNA product of 540 nt length. (B) A representative experiment as in A, but using the 19-nt LE19 RNA as a template. DN, PE, and TS indicate reaction products of de novo synthesis, primer extension, and template switching, respectively [ 54 ]. Procedures are detailed in Materials and Methods. Significance (Student’s T-test): ** p

    Article Snippet: To this end, 50 µl of 20 pmol/µl UTP, CTP, ATP and GTP (Jena Bioscience), were separated prior to sample analysis.

    Techniques: Activity Assay, Recombinant

    Effect of nucleoside-triphosphate concentration on NS5BΔ21 RNA polymerase activity. ( A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (100, 500, 800, and 1000 μM) of UTP in the presence of radiolabeled α 32 P-CTP. ATP and GTP concentrations were maintained at 100 μM and 500 μM, respectively (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ATP, with UTP and GTP maintained at 100 μM and 500 μM, respectively. (C) Corresponds to experiments as in A but for increasing concentrations of GTP, with ATP and UTP both maintained at 100 μM. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between the activity values that link, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): *** p

    Journal: bioRxiv

    Article Title: Guanosine inhibits hepatitis C virus replication and increases indel frequencies, associated with altered intracellular nucleotide pools

    doi: 10.1101/2020.02.21.959536

    Figure Lengend Snippet: Effect of nucleoside-triphosphate concentration on NS5BΔ21 RNA polymerase activity. ( A) Polyacrylamide gel showing the products for de novo (DN), primer extension (PE) and template switching (TS) obtained with HCV NS5BΔ21 at increasing concentrations (100, 500, 800, and 1000 μM) of UTP in the presence of radiolabeled α 32 P-CTP. ATP and GTP concentrations were maintained at 100 μM and 500 μM, respectively (left panel). Graphic representation of densitometric values obtained from the electropherogram shown in A (red diamonds, yellow triangles, and black squares correspond to de novo (DN), primer extension (PE), and template switching (TS) activities, respectively) (right panel). (B) Corresponds to experiments as in A but for increasing concentrations of ATP, with UTP and GTP maintained at 100 μM and 500 μM, respectively. (C) Corresponds to experiments as in A but for increasing concentrations of GTP, with ATP and UTP both maintained at 100 μM. Activities were normalized to their maximum values. Densitometric data represent the mean of at least three independent experiments. Error bars correspond to standard error of the mean. Horizontal lines indicate statistically significant differences (Student’s T-test) between the activity values that link, using the same color code as the activity type. Details of the activity measurements are given in Materials and Methods. Significance (Student’s T-test): *** p

    Article Snippet: To this end, 50 µl of 20 pmol/µl UTP, CTP, ATP and GTP (Jena Bioscience), were separated prior to sample analysis.

    Techniques: Concentration Assay, Activity Assay