dithiothreatol  (Thermo Fisher)


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    Structured Review

    Thermo Fisher dithiothreatol
    Dithiothreatol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dithiothreatol/product/Thermo Fisher
    Average 82 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dithiothreatol - by Bioz Stars, 2020-03
    82/100 stars

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    Related Articles

    Radioactivity:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: Paragraph title: Radiolabeling, Immunoprecipitation and Western Blotting ... The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

    Autoradiography:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies). .. Samples were separated on 8–16% Tris-acetate polyacrylamide gels (Invitrogen, Life Technologies), with equal amounts used for either immunoblot analysis or autoradiography.

    Immunoprecipitation:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: Paragraph title: Radiolabeling, Immunoprecipitation and Western Blotting ... The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

    Incubation:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: After separation on a magnetic stand (Promega, Madison WI), the antibody bound beads were washed once with PBS-T and the radiolabeled lysate added, followed by a 10 min incubation at room temperature. .. The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

    Protein Concentration:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: Lysates were adjusted to the same protein concentration with lysis buffer and 16.25 µl removed for immunoblotting. .. The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

    Western Blot:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: Paragraph title: Radiolabeling, Immunoprecipitation and Western Blotting ... The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

    Lysis:

    Article Title: Palmitoylation of the Immunity Related GTPase, Irgm1: Impact on Membrane Localization and Ability to Promote Mitochondrial Fission
    Article Snippet: Lysates were adjusted to the same protein concentration with lysis buffer and 16.25 µl removed for immunoblotting. .. The beads were washed 4 times with PBS-T and suspended in 30 µl 1× LDS sample buffer with 0.1 M dithiothreatol (Invitrogen, Life Technologies).

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  • 99
    Thermo Fisher dtt
    Dynamic range of <t>roGFP2-Orp1</t> and HyPer-3 in transfected parasites within intact RBCs and after host cell lysis. After 15 s baseline monitoring, 3D7 parasites with intact or lysed host cells and transfected with roGFP2-Orp1 ( A ) or HyPer-3 ( C ) were exposed to 1 mM DIA and monitored for 2 min before adding 10 mM <t>DTT</t> at the CLSM. The fluorescence ratios (405/488 nm, 3D7 [roGFP2-Orp1] and 488/405 nm, 3D7 [HyPer-3] ) ( A, C ) at different time points are plotted against time. 3D7 [HyPer-3] ( C ) showed a higher DIA sensitivity than 3D7 [roGFP2-Orp1] ( A ) in both parasites residing in intact RBCs and those deprived of their host cell. Data from at least three trophozoites in three independent experiments were analyzed per data point. For measuring the dynamic range of both redox sensors in the parasites, the 405/488 nm ratio (3D7 [roGFP2-Orp1] ) ( B ) and the 488/405 nm ratio (3D7 [HyPer-3] ) ( D ) of fully oxidized and reduced probes were computed. The basal ratio, the ratio for 1 mM DIA, and 10 mM DTT after 2 min incubation (n > 27) of 3 independent experiments are shown. 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) with intact RBCs exhibited dynamic ranges of 5 and 7.3, respectively. The dynamic ranges for parasites after RBC lysis of 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) were 5.5 and 12.6, respectively. Mean values and standard error of the mean (SEM) are shown for all experiments. A one-way ANOVA test with 95% confidence intervals with the Dunnett’s Multiple Comparison Test was applied for statistical analysis of significance (*, p
    Dtt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2501 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt/product/Thermo Fisher
    Average 99 stars, based on 2501 article reviews
    Price from $9.99 to $1999.99
    dtt - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    95
    Thermo Fisher m dithiothreitol
    Redox potential of the MetAP2 disulfide bond. A , the labeling of fully reduced MetAP2 by the biotin-linked maleimide, MPB, was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (DTT red ) to oxidized (DTT ox ) <t>dithiothreitol.</t> Biotin incorporation was measured by SDS-PAGE and blotting with streptavidin-peroxidase. B , plot of the ratio of reduced to oxidized MetAP2 as a function of the ratio of reduced to oxidized DTT. The solid line . The calculated equilibrium constant is 0.028 (95% confidence interval: 0.012–0.044). From the Nernst equation, the standard redox potential of the Cys 228 -Cys 448 disulfide is −261 mV. Data points and error bars , mean ± S.E. of three independent experiments. C , the initial rate of hydrolysis of Met-Gly-Pro-AMC by fully oxidized or reduced MetAP2 was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (Trx red ) to oxidized (Trx ox ) thioredoxin. The solid line . The calculated equilibrium constant is 0.740 (95% confidence interval: 0.600–0.688), which equates to a standard redox potential of −266 mV for the Cys 228 -Cys 448 disulfide bond. Data points and error bars , mean ± S.E. of three independent experiments.
    M Dithiothreitol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m dithiothreitol/product/Thermo Fisher
    Average 95 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    m dithiothreitol - by Bioz Stars, 2020-03
    95/100 stars
      Buy from Supplier

    Image Search Results


    Dynamic range of roGFP2-Orp1 and HyPer-3 in transfected parasites within intact RBCs and after host cell lysis. After 15 s baseline monitoring, 3D7 parasites with intact or lysed host cells and transfected with roGFP2-Orp1 ( A ) or HyPer-3 ( C ) were exposed to 1 mM DIA and monitored for 2 min before adding 10 mM DTT at the CLSM. The fluorescence ratios (405/488 nm, 3D7 [roGFP2-Orp1] and 488/405 nm, 3D7 [HyPer-3] ) ( A, C ) at different time points are plotted against time. 3D7 [HyPer-3] ( C ) showed a higher DIA sensitivity than 3D7 [roGFP2-Orp1] ( A ) in both parasites residing in intact RBCs and those deprived of their host cell. Data from at least three trophozoites in three independent experiments were analyzed per data point. For measuring the dynamic range of both redox sensors in the parasites, the 405/488 nm ratio (3D7 [roGFP2-Orp1] ) ( B ) and the 488/405 nm ratio (3D7 [HyPer-3] ) ( D ) of fully oxidized and reduced probes were computed. The basal ratio, the ratio for 1 mM DIA, and 10 mM DTT after 2 min incubation (n > 27) of 3 independent experiments are shown. 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) with intact RBCs exhibited dynamic ranges of 5 and 7.3, respectively. The dynamic ranges for parasites after RBC lysis of 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) were 5.5 and 12.6, respectively. Mean values and standard error of the mean (SEM) are shown for all experiments. A one-way ANOVA test with 95% confidence intervals with the Dunnett’s Multiple Comparison Test was applied for statistical analysis of significance (*, p

    Journal: PLoS ONE

    Article Title: H2O2 dynamics in the malaria parasite Plasmodium falciparum

    doi: 10.1371/journal.pone.0174837

    Figure Lengend Snippet: Dynamic range of roGFP2-Orp1 and HyPer-3 in transfected parasites within intact RBCs and after host cell lysis. After 15 s baseline monitoring, 3D7 parasites with intact or lysed host cells and transfected with roGFP2-Orp1 ( A ) or HyPer-3 ( C ) were exposed to 1 mM DIA and monitored for 2 min before adding 10 mM DTT at the CLSM. The fluorescence ratios (405/488 nm, 3D7 [roGFP2-Orp1] and 488/405 nm, 3D7 [HyPer-3] ) ( A, C ) at different time points are plotted against time. 3D7 [HyPer-3] ( C ) showed a higher DIA sensitivity than 3D7 [roGFP2-Orp1] ( A ) in both parasites residing in intact RBCs and those deprived of their host cell. Data from at least three trophozoites in three independent experiments were analyzed per data point. For measuring the dynamic range of both redox sensors in the parasites, the 405/488 nm ratio (3D7 [roGFP2-Orp1] ) ( B ) and the 488/405 nm ratio (3D7 [HyPer-3] ) ( D ) of fully oxidized and reduced probes were computed. The basal ratio, the ratio for 1 mM DIA, and 10 mM DTT after 2 min incubation (n > 27) of 3 independent experiments are shown. 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) with intact RBCs exhibited dynamic ranges of 5 and 7.3, respectively. The dynamic ranges for parasites after RBC lysis of 3D7 [roGFP2-Orp1] ( B ) and 3D7 [HyPer-3] ( D ) were 5.5 and 12.6, respectively. Mean values and standard error of the mean (SEM) are shown for all experiments. A one-way ANOVA test with 95% confidence intervals with the Dunnett’s Multiple Comparison Test was applied for statistical analysis of significance (*, p

    Article Snippet: Purified recombinant roGFP2-Orp1 and HyPer-3/SypHer proteins were reduced with 5 mM DTT for 10 min and 20 mM DTT for 30 min, respectively, at 4°C, desalinated (ZebaTM Spin Desalting Columns, Thermo Scientific), and diluted in reaction buffer to a final concentration of 5 μ M. A 5-fold drug/redox-active compound dilution (25 μ l) was mixed with 100 μ l of 5 μ M roGFP2-Orp1/HyPer in a 96-well microplate (black, half-area, Greiner Bio-One, Frickenhausen).

    Techniques: Transfection, Lysis, Confocal Laser Scanning Microscopy, Fluorescence, Incubation

    Dynamic range of roGFP2-Orp1 and Mito-roGFP2-Orp1 in transfected NF54- attB parasites. NF54- attB parasites transfected with roGFP2-Orp1 or Mito-roGFP2-Orp1 were exposed to 1 mM DIA or 10 mM DTT for 2 min before blocking with 2 mM NEM. Fluorescence ratios of 405/488 nm were detected with CLSM. NF54 [roGFP2-Orp1] - attB parasites showed a slightly higher DIA sensitivity than NF54 [Mito-roGFP2-Orp1] - attB parasites. CLSM data were composed of values from at least 10–20 trophozoites analyzed per experiment. Mean values and standard errors of the means (±SEM) are shown for three independent experiments. A one-way ANOVA test with 95% confidence intervals with the Dunnett’s Multiple Comparison Test was applied for statistical analysis of significance (***p

    Journal: Scientific Reports

    Article Title: Hydrogen peroxide dynamics in subcellular compartments of malaria parasites using genetically encoded redox probes

    doi: 10.1038/s41598-017-10093-8

    Figure Lengend Snippet: Dynamic range of roGFP2-Orp1 and Mito-roGFP2-Orp1 in transfected NF54- attB parasites. NF54- attB parasites transfected with roGFP2-Orp1 or Mito-roGFP2-Orp1 were exposed to 1 mM DIA or 10 mM DTT for 2 min before blocking with 2 mM NEM. Fluorescence ratios of 405/488 nm were detected with CLSM. NF54 [roGFP2-Orp1] - attB parasites showed a slightly higher DIA sensitivity than NF54 [Mito-roGFP2-Orp1] - attB parasites. CLSM data were composed of values from at least 10–20 trophozoites analyzed per experiment. Mean values and standard errors of the means (±SEM) are shown for three independent experiments. A one-way ANOVA test with 95% confidence intervals with the Dunnett’s Multiple Comparison Test was applied for statistical analysis of significance (***p

    Article Snippet: Purified recombinant roGFP2-Orp1 protein was reduced with 20 mM DTT for 30 min at 4 °C, desalinated (ZebaTM Spin Desalting Columns, Thermo Scientific), and diluted in reaction buffer to a final concentration of 5 µM.

    Techniques: Transfection, Blocking Assay, Fluorescence, Confocal Laser Scanning Microscopy

    Redox potential of the MetAP2 disulfide bond. A , the labeling of fully reduced MetAP2 by the biotin-linked maleimide, MPB, was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (DTT red ) to oxidized (DTT ox ) dithiothreitol. Biotin incorporation was measured by SDS-PAGE and blotting with streptavidin-peroxidase. B , plot of the ratio of reduced to oxidized MetAP2 as a function of the ratio of reduced to oxidized DTT. The solid line . The calculated equilibrium constant is 0.028 (95% confidence interval: 0.012–0.044). From the Nernst equation, the standard redox potential of the Cys 228 -Cys 448 disulfide is −261 mV. Data points and error bars , mean ± S.E. of three independent experiments. C , the initial rate of hydrolysis of Met-Gly-Pro-AMC by fully oxidized or reduced MetAP2 was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (Trx red ) to oxidized (Trx ox ) thioredoxin. The solid line . The calculated equilibrium constant is 0.740 (95% confidence interval: 0.600–0.688), which equates to a standard redox potential of −266 mV for the Cys 228 -Cys 448 disulfide bond. Data points and error bars , mean ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Redox Regulation of Methionine Aminopeptidase 2 Activity *

    doi: 10.1074/jbc.M114.554253

    Figure Lengend Snippet: Redox potential of the MetAP2 disulfide bond. A , the labeling of fully reduced MetAP2 by the biotin-linked maleimide, MPB, was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (DTT red ) to oxidized (DTT ox ) dithiothreitol. Biotin incorporation was measured by SDS-PAGE and blotting with streptavidin-peroxidase. B , plot of the ratio of reduced to oxidized MetAP2 as a function of the ratio of reduced to oxidized DTT. The solid line . The calculated equilibrium constant is 0.028 (95% confidence interval: 0.012–0.044). From the Nernst equation, the standard redox potential of the Cys 228 -Cys 448 disulfide is −261 mV. Data points and error bars , mean ± S.E. of three independent experiments. C , the initial rate of hydrolysis of Met-Gly-Pro-AMC by fully oxidized or reduced MetAP2 was used to calculate the ratio of reduced to oxidized enzyme as a function of the ratio of reduced (Trx red ) to oxidized (Trx ox ) thioredoxin. The solid line . The calculated equilibrium constant is 0.740 (95% confidence interval: 0.600–0.688), which equates to a standard redox potential of −266 mV for the Cys 228 -Cys 448 disulfide bond. Data points and error bars , mean ± S.E. of three independent experiments.

    Article Snippet: Just before use, the active site disulfide bond of the oxidoreductase was reduced with 50 m m dithiothreitol and 50 m m tris-(2-carboxyethyl)phosphine (TCEP) for 1 h at 25 °C, and the reducing agents were removed by two passes through Zeba spin desalting columns equilibrated with phosphate-buffered saline (Thermo Scientific).

    Techniques: Labeling, SDS Page