sonication dtt  (Thermo Fisher)


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    Structured Review

    Thermo Fisher sonication dtt
    Sonication Dtt, supplied by Thermo Fisher, 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/sonication dtt/product/Thermo Fisher
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sonication dtt - by Bioz Stars, 2020-03
    80/100 stars

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    Sample Prep:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: Paragraph title: Sample preparation ... Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris.

    Homogenization:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: .. Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Size-exclusion Chromatography:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: Samples were then carefully sonicated at 4°C for 15 sec (3 bursts for 5 sec) with Bandelin sonicator at 50% power output. .. Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris.

    Electrophoresis:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Concentration Assay:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: .. Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Incubation:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: .. Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Protein Concentration:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Sonication:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: .. Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

    Staining:

    Article Title: Integrated network analysis of transcriptomic and proteomic data in psoriasis
    Article Snippet: Following homogenization and sonication DTT (Acros) was added to reach the final concentration of 65 mM, and samples left at 4°C for 30 min. Then Na2 EDTA was added (2 mM final) and mixture was incubated for additional 2 h and then centrifuged at 12,000 × g for 10 min to remove insoluble debris. .. Relative protein concentration was determined with GS-800 Calibrated Densitometer (Bio-Rad) after 1D-SDS-PAGE of 3 μl sample solution loaded on 8 × 9 cm mini-gel stained with Coomassie R-250 after electrophoresis.

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    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
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    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