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  • 95
    Millipore datp
    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM <t>ATP</t> or <t>dATP.</t> Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.
    Datp, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore deoxynucleotide
    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM <t>ATP</t> or <t>dATP.</t> Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.
    Deoxynucleotide, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore deoxyadenosine triphosphate
    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM <t>ATP</t> or <t>dATP.</t> Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.
    Deoxyadenosine Triphosphate, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore c10 datp
    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM <t>ATP</t> or <t>dATP.</t> Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.
    C10 Datp, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore α 32 p datp labelled atal7 specific probes
    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM <t>ATP</t> or <t>dATP.</t> Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.
    α 32 P Datp Labelled Atal7 Specific Probes, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore 2 deoxy atp
    A–C : effect of <t>2-deoxy</t> <t>ATP</t> (dATP) of the velocity-pCa relationship at different pH. V RTF with WT Tn plotted as a function of the pCa and pH in the presence of dATP. Data points represent means ± SE. Data were fit with Hill equation with
    2 Deoxy Atp, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore biotin 14 datp
    A–C : effect of <t>2-deoxy</t> <t>ATP</t> (dATP) of the velocity-pCa relationship at different pH. V RTF with WT Tn plotted as a function of the pCa and pH in the presence of dATP. Data points represent means ± SE. Data were fit with Hill equation with
    Biotin 14 Datp, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dapt
    A–C : effect of <t>2-deoxy</t> <t>ATP</t> (dATP) of the velocity-pCa relationship at different pH. V RTF with WT Tn plotted as a function of the pCa and pH in the presence of dATP. Data points represent means ± SE. Data were fit with Hill equation with
    Dapt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.

    Journal: Nucleic Acids Research

    Article Title: Bacillus subtilis RecO and SsbA are crucial for RecA-mediated recombinational DNA repair

    doi: 10.1093/nar/gkv545

    Figure Lengend Snippet: Effect of RecO on RecA nucleation and polymerization. Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with the indicated concentration of RecO for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results. The amount of ATP or dATP hydrolyzed was calculated as described in ‘Materials and Methods’ section.

    Article Snippet: IPTG was from Calbiochem; DNA restriction enzymes, DNA ligase, etc. were supplied by Roche; and polyethyleneimine, DTT, ATP, dATP, ATPγS and AMP–PNP were from Sigma.

    Techniques: Incubation, Concentration Assay, Activity Assay

    RecO facilitates RecA assembly on SsbA- or SsbB-coated ssDNA. ( A ) Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with SsbA (0.3 μM) or SsbA and RecO for 5 min at 37°C in buffer A containing 5 mM ATP. Then RecA (0.8 μM) was added and the ATPase activity measured for 25 min. ( B ) Circular ssDNA was pre-incubated with SsbB (0.3 μM) (or both SsbA [0.3 μM] and SsbB [0.3 μM]) and when indicated with RecO (0.1 and 0.2 μM) for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. ( C ) The circular ssDNA was pre-incubated with a fixed amount of SsbB (0.6 μM), SsbA (0.3 μM) plus SsbB (0.3 μM) or a fixed amount of SsbB (0.3 μM) and increasing SsbA (0.03 to 0.3 μM) concentrations for 5 min at 37°C in buffer A containing 5 mM ATP. Then RecO (0.1 μM) was added to the preformed SSB·ssDNA complexes and incubated for 5 min at 37°C. Finally RecA (0.8 μM) was added and the ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results.

    Journal: Nucleic Acids Research

    Article Title: Bacillus subtilis RecO and SsbA are crucial for RecA-mediated recombinational DNA repair

    doi: 10.1093/nar/gkv545

    Figure Lengend Snippet: RecO facilitates RecA assembly on SsbA- or SsbB-coated ssDNA. ( A ) Circular pGEM3 Zf (+) ssDNA (10 μM in nt) was pre-incubated with SsbA (0.3 μM) or SsbA and RecO for 5 min at 37°C in buffer A containing 5 mM ATP. Then RecA (0.8 μM) was added and the ATPase activity measured for 25 min. ( B ) Circular ssDNA was pre-incubated with SsbB (0.3 μM) (or both SsbA [0.3 μM] and SsbB [0.3 μM]) and when indicated with RecO (0.1 and 0.2 μM) for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. Then RecA (0.8 μM) was added and the (d)ATPase activity measured for 25 min. ( C ) The circular ssDNA was pre-incubated with a fixed amount of SsbB (0.6 μM), SsbA (0.3 μM) plus SsbB (0.3 μM) or a fixed amount of SsbB (0.3 μM) and increasing SsbA (0.03 to 0.3 μM) concentrations for 5 min at 37°C in buffer A containing 5 mM ATP. Then RecO (0.1 μM) was added to the preformed SSB·ssDNA complexes and incubated for 5 min at 37°C. Finally RecA (0.8 μM) was added and the ATPase activity measured for 25 min. All reactions were repeated three or more times with similar results.

    Article Snippet: IPTG was from Calbiochem; DNA restriction enzymes, DNA ligase, etc. were supplied by Roche; and polyethyleneimine, DTT, ATP, dATP, ATPγS and AMP–PNP were from Sigma.

    Techniques: Incubation, Activity Assay

    SsbA and RecO contribute to RecA-promoted DNA strand exchange. ( A ) Circular pGEM3 Zf (+) ssDNA (10 μM in nt) and homologous linear dsDNA (20 μM in nt) were pre-incubated with increasing concentrations of SsbA or SsbB (0.15 and 0.3 μM) or RecO (0.2 and 0.4 μM) for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. In the indicated reactions, RecO (0.2 and 0.4 μM) was added to the preformed SsbA·ssDNA or SsbB·ssDNA (SSB 0.3 μM) complexes and incubated for 5 min at 37°C. Finally, RecA (1.2 μM) was added and the reaction was incubated for 60 min at 37°C. ( B ) The circular ssDNA and homologous linear dsDNA were pre-incubated with SsbA or SsbB (0.3 μM) for 5 min at 37°C in buffer A containing 5 mM dATP, ATP or ATPγS, and RecO (0.2 μM) was added to the preformed SsbA·ssDNA or SsbB·ssDNA complexes and incubated for 10 min at 37°C. Finally, RecA (1.2 μM) was added and the reaction was incubated for 60 min at 37°C. The products of the reactions were deproteinized, separated on a 0.8% AGE with ethidium bromide and quantified as described in ‘Materials and Methods’ section. The positions of the bands corresponding to css, cds, lds, jm, nc and ATPγS-generated prd products are indicated. Symbols + and − denote the presence or absence, respectively, of the indicated protein. In A (lane 14) and in B (lane 17) nicked circular pGEM3 Zf (+) plasmid DNA was added as a mobility control ( C ). The amounts of jm s and products ( nc and prd ) are indicated and expressed as the percentage of total substrate added. The results are the average value obtained from more than three independent experiments (the results given stand within a 5% standard error).

    Journal: Nucleic Acids Research

    Article Title: Bacillus subtilis RecO and SsbA are crucial for RecA-mediated recombinational DNA repair

    doi: 10.1093/nar/gkv545

    Figure Lengend Snippet: SsbA and RecO contribute to RecA-promoted DNA strand exchange. ( A ) Circular pGEM3 Zf (+) ssDNA (10 μM in nt) and homologous linear dsDNA (20 μM in nt) were pre-incubated with increasing concentrations of SsbA or SsbB (0.15 and 0.3 μM) or RecO (0.2 and 0.4 μM) for 5 min at 37°C in buffer A containing 5 mM ATP or dATP. In the indicated reactions, RecO (0.2 and 0.4 μM) was added to the preformed SsbA·ssDNA or SsbB·ssDNA (SSB 0.3 μM) complexes and incubated for 5 min at 37°C. Finally, RecA (1.2 μM) was added and the reaction was incubated for 60 min at 37°C. ( B ) The circular ssDNA and homologous linear dsDNA were pre-incubated with SsbA or SsbB (0.3 μM) for 5 min at 37°C in buffer A containing 5 mM dATP, ATP or ATPγS, and RecO (0.2 μM) was added to the preformed SsbA·ssDNA or SsbB·ssDNA complexes and incubated for 10 min at 37°C. Finally, RecA (1.2 μM) was added and the reaction was incubated for 60 min at 37°C. The products of the reactions were deproteinized, separated on a 0.8% AGE with ethidium bromide and quantified as described in ‘Materials and Methods’ section. The positions of the bands corresponding to css, cds, lds, jm, nc and ATPγS-generated prd products are indicated. Symbols + and − denote the presence or absence, respectively, of the indicated protein. In A (lane 14) and in B (lane 17) nicked circular pGEM3 Zf (+) plasmid DNA was added as a mobility control ( C ). The amounts of jm s and products ( nc and prd ) are indicated and expressed as the percentage of total substrate added. The results are the average value obtained from more than three independent experiments (the results given stand within a 5% standard error).

    Article Snippet: IPTG was from Calbiochem; DNA restriction enzymes, DNA ligase, etc. were supplied by Roche; and polyethyleneimine, DTT, ATP, dATP, ATPγS and AMP–PNP were from Sigma.

    Techniques: Incubation, Generated, Plasmid Preparation

    Representative chromatogram of dNTP levels at the LLOQ Ion chromatograms showing relative abundance for dGTP, ATP, dATP, labeled C-13 dATP, dTTP, and dCTP upon injection of 62.5 fmol dNTPs, 1,250 fmol ATP, and 2.5 pmol of labeled C-13 dATP.

    Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

    Article Title: Fast and sensitive HPLC-MS/MS method for direct quantification of intracellular deoxyribonucleoside triphosphates from tissue and cells

    doi: 10.1016/j.jchromb.2017.10.008

    Figure Lengend Snippet: Representative chromatogram of dNTP levels at the LLOQ Ion chromatograms showing relative abundance for dGTP, ATP, dATP, labeled C-13 dATP, dTTP, and dCTP upon injection of 62.5 fmol dNTPs, 1,250 fmol ATP, and 2.5 pmol of labeled C-13 dATP.

    Article Snippet: The internal standard used was C-13 labeled dATP, purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Labeling, Injection

    A–C : effect of 2-deoxy ATP (dATP) of the velocity-pCa relationship at different pH. V RTF with WT Tn plotted as a function of the pCa and pH in the presence of dATP. Data points represent means ± SE. Data were fit with Hill equation with

    Journal: Journal of Applied Physiology

    Article Title: Ca++-sensitizing mutations in troponin, Pi, and 2-deoxyATP alter the depressive effect of acidosis on regulated thin-filament velocity

    doi: 10.1152/japplphysiol.01161.2013

    Figure Lengend Snippet: A–C : effect of 2-deoxy ATP (dATP) of the velocity-pCa relationship at different pH. V RTF with WT Tn plotted as a function of the pCa and pH in the presence of dATP. Data points represent means ± SE. Data were fit with Hill equation with

    Article Snippet: The buffers for the motility assay were based on a lower-salt actin buffer (25 mM KCl, 25 mM imidazole, 1 mM EGTA, 4 mM MgCl2 , 1 mM DTT) and 2 mM ATP or 2-deoxy-ATP (Sigma-Aldrich).

    Techniques:

    Effect of deoxy-ATP (dATP) on maximal force ( F max , pCa 4.0) in skeletal ( A ) and cardiac muscle ( B ) with intact and isolated regulatory units (RUs)

    Journal:

    Article Title: Investigation of thin filament near-neighbour regulatory unit interactions during force development in skinned cardiac and skeleta muscle

    doi: 10.1113/jphysiol.2007.128975

    Figure Lengend Snippet: Effect of deoxy-ATP (dATP) on maximal force ( F max , pCa 4.0) in skeletal ( A ) and cardiac muscle ( B ) with intact and isolated regulatory units (RUs)

    Article Snippet: Solutions contained (m m ): phosphocreatine 15, EGTA 15, Mops at least 40, free Mg2+ 1, Na+ + K+ 135 and DTT 1, 250 units ml−1 creatine kinase (Sigma) and 5 m m ATP, 0.5 m m ATP, or 5 m m 2 deoxy-ATP (dATP; Sigma) at pH 7.0 and 15 ± 1°C.

    Techniques: Isolation

    Stimulation of open probability by 2'-deoxy-ATP (dATP) or N 6 -(2-phenylethyl)-dATP (P-dATP) facilitates counting channels for low-P o mutants. Inward currents of single cut-ΔR(D1370N) channels harbouring the indicated mutations. After several minutes of low-P o activity in 10 mM MgATP ( green bars ), exposure to 5 mM Mg-dATP or 25–50 μM Mg-P-dATP ( red bars ) robustly stimulated P o . A statistical test based on comparison of the observed opening rate and cumulative open time ( Csanády et al., 2000 ) allowed high-confidence exclusion of the presence of a second active channel for each of the four patches shown (p

    Journal: eLife

    Article Title: Asymmetry of movements in CFTR's two ATP sites during pore opening serves their distinct functions

    doi: 10.7554/eLife.29013

    Figure Lengend Snippet: Stimulation of open probability by 2'-deoxy-ATP (dATP) or N 6 -(2-phenylethyl)-dATP (P-dATP) facilitates counting channels for low-P o mutants. Inward currents of single cut-ΔR(D1370N) channels harbouring the indicated mutations. After several minutes of low-P o activity in 10 mM MgATP ( green bars ), exposure to 5 mM Mg-dATP or 25–50 μM Mg-P-dATP ( red bars ) robustly stimulated P o . A statistical test based on comparison of the observed opening rate and cumulative open time ( Csanády et al., 2000 ) allowed high-confidence exclusion of the presence of a second active channel for each of the four patches shown (p

    Article Snippet: The bath solution was continuously flowing, and could be exchanged with a time constant of ~100 ms. MgATP (3 or 10 mM) and 2'-deoxy-ATP sodium salt (dATP, 5 mM) (Sigma) were added from 400 mM and 100 mM aqueous stock solutions, respectively (pH = 7.1 with NMDG); dATP was supplemented with equimolar MgCl2 .

    Techniques: Activity Assay