ctp  (Jena Bioscience)


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  • 93
    Name:
    CTP Solid
    Description:

    Catalog Number:
    NU-1011-100
    Price:
    50.4
    Category:
    Molecular Biology
    Size:
    100 mg
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    Structured Review

    Jena Bioscience ctp

    https://www.bioz.com/result/ctp/product/Jena Bioscience
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ctp - by Bioz Stars, 2021-07
    93/100 stars

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    Injection:

    Article Title: Structural and biochemical insights into inhibition of human primase by citrate.
    Article Snippet: The eukaryotic primase/polymerase complex synthesizes approximately 10 < sup > 7 < /sup > primers, one per Okazaki fragment, during the replication of mammalian chromosomes, which contain 10 < sup > 9 < /sup > base pairs. .. The eukaryotic primase/polymerase complex synthesizes approximately 10 < sup > 7 < /sup > primers, one per Okazaki fragment, during the replication of mammalian chromosomes, which contain 10 < sup > 9 < /sup > base pairs. ..

    Purification:

    Article Title: Structural and biochemical insights into inhibition of human primase by citrate.
    Article Snippet: The eukaryotic primase/polymerase complex synthesizes approximately 10 < sup > 7 < /sup > primers, one per Okazaki fragment, during the replication of mammalian chromosomes, which contain 10 < sup > 9 < /sup > base pairs. .. The eukaryotic primase/polymerase complex synthesizes approximately 10 < sup > 7 < /sup > primers, one per Okazaki fragment, during the replication of mammalian chromosomes, which contain 10 < sup > 9 < /sup > base pairs. ..

    Nuclear Magnetic Resonance:

    Article Title: Time-resolved NMR monitoring of tRNA maturation
    Article Snippet: All strain constructions were verified by PCR using appropriate oligonucleotides (listed in Supplementary Table ). .. tRNAPhe samples for NMR Unmodified yeast tRNAPhe was prepared by standard in vitro transcription with T7 polymerase with unlabeled NTPs (Jena Bioscience) for unlabeled samples or 15 N-labeled UTP and GTP (Eurisotop) and unlabeled ATP and CTP (Jena Bioscience) for 15 N-[U/G]-labeled samples. .. The DNA template and T7 promotor primer were purchased from Eurogentec.

    In Vitro:

    Article Title: Time-resolved NMR monitoring of tRNA maturation
    Article Snippet: All strain constructions were verified by PCR using appropriate oligonucleotides (listed in Supplementary Table ). .. tRNAPhe samples for NMR Unmodified yeast tRNAPhe was prepared by standard in vitro transcription with T7 polymerase with unlabeled NTPs (Jena Bioscience) for unlabeled samples or 15 N-labeled UTP and GTP (Eurisotop) and unlabeled ATP and CTP (Jena Bioscience) for 15 N-[U/G]-labeled samples. .. The DNA template and T7 promotor primer were purchased from Eurogentec.

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  • 93
    Jena Bioscience ara ctp
    The kinetic and morphological comparison of the nuclear envelope breakdown between replication-completed nuclei, nuclei with stalled forks, and nuclei with released and naturally ongoing forks . A , experimental strategy. Without replication stress, sperm nuclei generally replicate their genomic DNA in 30 to 60 min in the S-phase egg extract. Therefore, there are naturally ongoing forks at 35 min and no forks at 80 min. Stalled forks are stabilized at 80 min in the presence of <t>Ara-CTP.</t> Then, mitotic entry was induced in the presence or absence of Ara-CTP or <t>ex-dCTP,</t> and nuclei were fixed at each time point and observed by microscopy. B , the time courses of the NEB rates were determined as in Figure 3 . The same experiments were repeated three times. Error bar, ± SD. C , the pictures of nuclei representative at each time point. The genomic DNA and nuclear envelope were stained with Hoechst 33258 and DiOC6, respectively. Bar, 20 μm. Ara-CTP, Ara-cytidine-5′-triphosphate; DiOC6, 3,3′-dihexyloxacarbocyanine iodide.
    Ara Ctp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ara ctp/product/Jena Bioscience
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ara ctp - by Bioz Stars, 2021-07
    93/100 stars
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    The kinetic and morphological comparison of the nuclear envelope breakdown between replication-completed nuclei, nuclei with stalled forks, and nuclei with released and naturally ongoing forks . A , experimental strategy. Without replication stress, sperm nuclei generally replicate their genomic DNA in 30 to 60 min in the S-phase egg extract. Therefore, there are naturally ongoing forks at 35 min and no forks at 80 min. Stalled forks are stabilized at 80 min in the presence of Ara-CTP. Then, mitotic entry was induced in the presence or absence of Ara-CTP or ex-dCTP, and nuclei were fixed at each time point and observed by microscopy. B , the time courses of the NEB rates were determined as in Figure 3 . The same experiments were repeated three times. Error bar, ± SD. C , the pictures of nuclei representative at each time point. The genomic DNA and nuclear envelope were stained with Hoechst 33258 and DiOC6, respectively. Bar, 20 μm. Ara-CTP, Ara-cytidine-5′-triphosphate; DiOC6, 3,3′-dihexyloxacarbocyanine iodide.

    Journal: The Journal of Biological Chemistry

    Article Title: Ongoing replication forks delay the nuclear envelope breakdown upon mitotic entry

    doi: 10.1074/jbc.RA120.015142

    Figure Lengend Snippet: The kinetic and morphological comparison of the nuclear envelope breakdown between replication-completed nuclei, nuclei with stalled forks, and nuclei with released and naturally ongoing forks . A , experimental strategy. Without replication stress, sperm nuclei generally replicate their genomic DNA in 30 to 60 min in the S-phase egg extract. Therefore, there are naturally ongoing forks at 35 min and no forks at 80 min. Stalled forks are stabilized at 80 min in the presence of Ara-CTP. Then, mitotic entry was induced in the presence or absence of Ara-CTP or ex-dCTP, and nuclei were fixed at each time point and observed by microscopy. B , the time courses of the NEB rates were determined as in Figure 3 . The same experiments were repeated three times. Error bar, ± SD. C , the pictures of nuclei representative at each time point. The genomic DNA and nuclear envelope were stained with Hoechst 33258 and DiOC6, respectively. Bar, 20 μm. Ara-CTP, Ara-cytidine-5′-triphosphate; DiOC6, 3,3′-dihexyloxacarbocyanine iodide.

    Article Snippet: Therefore, the final concentrations of Ara-CTP and excess dCTP were 100 μM and 2 mM in the presence of the M-extract, and 200 μM and 4 mM in the absence of the M-extract, respectively, except that 200 μM of Ara-CTP and 4 mM of excess dCTP in the presence of M-extract in ( ).

    Techniques: Acetylene Reduction Assay, Microscopy, Staining

    Replication forks arrested by Ara-CTP are kept competent to restart in the S phase . Without excess dCTP, mitotic entry drives the nuclear envelope breakdown (NEB) and replisome disassembly, rendering stalled forks unable to restart. In contrast, DNA replication efficiently resumes with excess dCTP while the NEB is delayed in a Wee1/Myt1–dependent manner. It is unclear whether released forks accomplish DNA replication or are subjected to mitotic replisome disassembly on the way. Ara-CTP, Ara-cytidine-5′-triphosphate.

    Journal: The Journal of Biological Chemistry

    Article Title: Ongoing replication forks delay the nuclear envelope breakdown upon mitotic entry

    doi: 10.1074/jbc.RA120.015142

    Figure Lengend Snippet: Replication forks arrested by Ara-CTP are kept competent to restart in the S phase . Without excess dCTP, mitotic entry drives the nuclear envelope breakdown (NEB) and replisome disassembly, rendering stalled forks unable to restart. In contrast, DNA replication efficiently resumes with excess dCTP while the NEB is delayed in a Wee1/Myt1–dependent manner. It is unclear whether released forks accomplish DNA replication or are subjected to mitotic replisome disassembly on the way. Ara-CTP, Ara-cytidine-5′-triphosphate.

    Article Snippet: Therefore, the final concentrations of Ara-CTP and excess dCTP were 100 μM and 2 mM in the presence of the M-extract, and 200 μM and 4 mM in the absence of the M-extract, respectively, except that 200 μM of Ara-CTP and 4 mM of excess dCTP in the presence of M-extract in ( ).

    Techniques: Acetylene Reduction Assay

    Stalled replication forks can restart during early mitosis when replication stress is relieved . A , experimental strategy. B , in the first reaction, sperm nuclei were incubated in an S-phase extract (S-extract) with Ara-CTP for 60 min. In the second reaction, an equal volume of the M-phase extract (M-extract) containing Cy5-dUTP and Ara-CTP or excess dCTP (ex-dCTP) was added to the first reaction mixture and further incubated for 30 to 60 min. Then, genomic DNA was isolated and subjected to 0.8% TAE agarose gel electrophoresis, followed by SYBR Gold staining. Detected fluorescent signals of Cy5 and SYBR Gold show replicated DNA and total DNA, respectively. Recombinant His-p27 (p27) was added to inhibit CDK activities and mitotic entry. C , the same experiment in panel B was repeated three times, and the signal intensities of Cy5 were quantified by ImageJ. The replication activity obtained after 90 min incubation in the S-phase extract under unperturbed condition was used as a control for normalization. The relative replication activities were plotted in the graph. Error bar, ± SD. D , the same experiment in panel B was performed using CF594-dUTP instead of Cy5-dUTP to detect replication activity. After 30 min in the second reaction, nuclei were fixed and observed by fluorescence microscopy. Nuclear DNA was stained with Hoechst 33258. Bar, 20 μm. E – F , replication activities at each time point were monitored by pulse labeling with Cy5-dUTP added 6 min before isolating genomic DNA, which was analyzed in a similar way as shown in panels B – C . The replication product at 10 min in the presence of ex-dCTP and p27 was used as the control. Ara-CTP, Ara-cytidine-5′-triphosphate; CDK, cyclin-dependent kinase.

    Journal: The Journal of Biological Chemistry

    Article Title: Ongoing replication forks delay the nuclear envelope breakdown upon mitotic entry

    doi: 10.1074/jbc.RA120.015142

    Figure Lengend Snippet: Stalled replication forks can restart during early mitosis when replication stress is relieved . A , experimental strategy. B , in the first reaction, sperm nuclei were incubated in an S-phase extract (S-extract) with Ara-CTP for 60 min. In the second reaction, an equal volume of the M-phase extract (M-extract) containing Cy5-dUTP and Ara-CTP or excess dCTP (ex-dCTP) was added to the first reaction mixture and further incubated for 30 to 60 min. Then, genomic DNA was isolated and subjected to 0.8% TAE agarose gel electrophoresis, followed by SYBR Gold staining. Detected fluorescent signals of Cy5 and SYBR Gold show replicated DNA and total DNA, respectively. Recombinant His-p27 (p27) was added to inhibit CDK activities and mitotic entry. C , the same experiment in panel B was repeated three times, and the signal intensities of Cy5 were quantified by ImageJ. The replication activity obtained after 90 min incubation in the S-phase extract under unperturbed condition was used as a control for normalization. The relative replication activities were plotted in the graph. Error bar, ± SD. D , the same experiment in panel B was performed using CF594-dUTP instead of Cy5-dUTP to detect replication activity. After 30 min in the second reaction, nuclei were fixed and observed by fluorescence microscopy. Nuclear DNA was stained with Hoechst 33258. Bar, 20 μm. E – F , replication activities at each time point were monitored by pulse labeling with Cy5-dUTP added 6 min before isolating genomic DNA, which was analyzed in a similar way as shown in panels B – C . The replication product at 10 min in the presence of ex-dCTP and p27 was used as the control. Ara-CTP, Ara-cytidine-5′-triphosphate; CDK, cyclin-dependent kinase.

    Article Snippet: Therefore, the final concentrations of Ara-CTP and excess dCTP were 100 μM and 2 mM in the presence of the M-extract, and 200 μM and 4 mM in the absence of the M-extract, respectively, except that 200 μM of Ara-CTP and 4 mM of excess dCTP in the presence of M-extract in ( ).

    Techniques: Incubation, Acetylene Reduction Assay, Isolation, Agarose Gel Electrophoresis, Staining, Recombinant, Activity Assay, Fluorescence, Microscopy, Labeling

    Stalled replication forks fail to restart after the nuclear envelope breakdown . A , experimental strategy. B – D , after the first reaction, an M-phase extract and Ara-CTP were added to the reaction mixture and then equally divided into four ( B and D ) or three pieces ( C ), to each of which excess amount of dCTP (ex-dCTP, dC) was added at 0, 10, 20, and 30 min ( B and D ) (at 0′, at 10′, at 20′, at 30′) or at 0 and 15 min ( C ) (at 0′ and at 15′) in the second reaction. B , the time courses of the average NEB rates of each sample were measured as in Figure 3 and shown in the graph. The same experiments were repeated three times. Error bar, ± SD. C , nuclear fractions were isolated at each time point and subjected to immunoblotting. The samples without excess dCTP addition were also prepared (−dC). D , in the second reaction, Cy5-dUTP was added at the same time with ex-dCTP and further incubated for 15 min, and the genomic DNA was isolated and subjected to 0.8% TAE agarose gel electrophoresis. The same experiments were repeated three times, and the relative replication activities are plotted in the graph in panel E together with the NEB rate at the time when ex-dCTP was added. Error bar, ± SD. Ara-CTP, Ara-cytidine-5′-triphosphate.

    Journal: The Journal of Biological Chemistry

    Article Title: Ongoing replication forks delay the nuclear envelope breakdown upon mitotic entry

    doi: 10.1074/jbc.RA120.015142

    Figure Lengend Snippet: Stalled replication forks fail to restart after the nuclear envelope breakdown . A , experimental strategy. B – D , after the first reaction, an M-phase extract and Ara-CTP were added to the reaction mixture and then equally divided into four ( B and D ) or three pieces ( C ), to each of which excess amount of dCTP (ex-dCTP, dC) was added at 0, 10, 20, and 30 min ( B and D ) (at 0′, at 10′, at 20′, at 30′) or at 0 and 15 min ( C ) (at 0′ and at 15′) in the second reaction. B , the time courses of the average NEB rates of each sample were measured as in Figure 3 and shown in the graph. The same experiments were repeated three times. Error bar, ± SD. C , nuclear fractions were isolated at each time point and subjected to immunoblotting. The samples without excess dCTP addition were also prepared (−dC). D , in the second reaction, Cy5-dUTP was added at the same time with ex-dCTP and further incubated for 15 min, and the genomic DNA was isolated and subjected to 0.8% TAE agarose gel electrophoresis. The same experiments were repeated three times, and the relative replication activities are plotted in the graph in panel E together with the NEB rate at the time when ex-dCTP was added. Error bar, ± SD. Ara-CTP, Ara-cytidine-5′-triphosphate.

    Article Snippet: Therefore, the final concentrations of Ara-CTP and excess dCTP were 100 μM and 2 mM in the presence of the M-extract, and 200 μM and 4 mM in the absence of the M-extract, respectively, except that 200 μM of Ara-CTP and 4 mM of excess dCTP in the presence of M-extract in ( ).

    Techniques: Acetylene Reduction Assay, Isolation, Incubation, Agarose Gel Electrophoresis

    Knockout of DCK using a CRISPR‐Cas9 system in KOPN41 cells. (A) Schematic representation of the target sequence of CRISPR‐Cas9. (B) Sequence of the cleavage site in clones #1 and #2. Boxes indicate identical sequences. (C) DCK protein expression in knockout clones and wild‐type (WT) cells as determined by immunoblotting. (D) Intracellular concentrations of Ara‐CTP in wild‐type and DCK knockout (KO) clone of KOPN41. Cells were cultured in the absence or presence of 0.8 (195), 4 (973), and 20 μ mol/L (4864 ng/mL) of Ara‐C for 6 h, and intracellular Ara‐CTP was measured by liquid chromatography–mass spectrometry (LC/MS). The vertical axis indicates intracellular concentration of Ara‐CTP (pmol/1.5 × 10 7 cells), and the horizontal axis indicates the concentration of Ara‐C ( μ mol/L). Error bars indicate standard error of triplicate experiment. Asterisks indicate significance (* P

    Journal: Cancer Medicine

    Article Title: Clofarabine exerts antileukemic activity against cytarabine‐resistant B‐cell precursor acute lymphoblastic leukemia with low deoxycytidine kinase expression

    doi: 10.1002/cam4.1323

    Figure Lengend Snippet: Knockout of DCK using a CRISPR‐Cas9 system in KOPN41 cells. (A) Schematic representation of the target sequence of CRISPR‐Cas9. (B) Sequence of the cleavage site in clones #1 and #2. Boxes indicate identical sequences. (C) DCK protein expression in knockout clones and wild‐type (WT) cells as determined by immunoblotting. (D) Intracellular concentrations of Ara‐CTP in wild‐type and DCK knockout (KO) clone of KOPN41. Cells were cultured in the absence or presence of 0.8 (195), 4 (973), and 20 μ mol/L (4864 ng/mL) of Ara‐C for 6 h, and intracellular Ara‐CTP was measured by liquid chromatography–mass spectrometry (LC/MS). The vertical axis indicates intracellular concentration of Ara‐CTP (pmol/1.5 × 10 7 cells), and the horizontal axis indicates the concentration of Ara‐C ( μ mol/L). Error bars indicate standard error of triplicate experiment. Asterisks indicate significance (* P

    Article Snippet: We next measured intracellular concentration of Ara‐CTP in both wild‐type and DCK knockout clone (#1) of KOPN41, which were cultured in the absence or presence of 0.8–20 μ mol/L (195–4864 ng/mL) of Ara‐C for 6 h, using liquid chromatography–mass spectrometry (LC/MS).

    Techniques: Knock-Out, CRISPR, Sequencing, Clone Assay, Expressing, Acetylene Reduction Assay, Cell Culture, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Concentration Assay

    Knockout of DCK using a CRISPR‐Cas9 system in KOPN41 cells. (A) Schematic representation of the target sequence of CRISPR‐Cas9. (B) Sequence of the cleavage site in clones #1 and #2. Boxes indicate identical sequences. (C) DCK protein expression in knockout clones and wild‐type (WT) cells as determined by immunoblotting. (D) Intracellular concentrations of Ara‐CTP in wild‐type and DCK knockout (KO) clone of KOPN41. Cells were cultured in the absence or presence of 0.8 (195), 4 (973), and 20 μ mol/L (4864 ng/mL) of Ara‐C for 6 h, and intracellular Ara‐CTP was measured by liquid chromatography–mass spectrometry (LC/MS). The vertical axis indicates intracellular concentration of Ara‐CTP (pmol/1.5 × 10 7 cells), and the horizontal axis indicates the concentration of Ara‐C ( μ mol/L). Error bars indicate standard error of triplicate experiment. Asterisks indicate significance (* P

    Journal: Cancer Medicine

    Article Title: Clofarabine exerts antileukemic activity against cytarabine‐resistant B‐cell precursor acute lymphoblastic leukemia with low deoxycytidine kinase expression

    doi: 10.1002/cam4.1323

    Figure Lengend Snippet: Knockout of DCK using a CRISPR‐Cas9 system in KOPN41 cells. (A) Schematic representation of the target sequence of CRISPR‐Cas9. (B) Sequence of the cleavage site in clones #1 and #2. Boxes indicate identical sequences. (C) DCK protein expression in knockout clones and wild‐type (WT) cells as determined by immunoblotting. (D) Intracellular concentrations of Ara‐CTP in wild‐type and DCK knockout (KO) clone of KOPN41. Cells were cultured in the absence or presence of 0.8 (195), 4 (973), and 20 μ mol/L (4864 ng/mL) of Ara‐C for 6 h, and intracellular Ara‐CTP was measured by liquid chromatography–mass spectrometry (LC/MS). The vertical axis indicates intracellular concentration of Ara‐CTP (pmol/1.5 × 10 7 cells), and the horizontal axis indicates the concentration of Ara‐C ( μ mol/L). Error bars indicate standard error of triplicate experiment. Asterisks indicate significance (* P

    Article Snippet: We next measured intracellular concentration of Ara‐CTP in both wild‐type and DCK knockout clone (#1) of KOPN41, which were cultured in the absence or presence of 0.8–20 μ mol/L (195–4864 ng/mL) of Ara‐C for 6 h, using liquid chromatography–mass spectrometry (LC/MS).

    Techniques: Knock-Out, CRISPR, Sequencing, Clone Assay, Expressing, Acetylene Reduction Assay, Cell Culture, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Concentration Assay