dnase i  (Worthington Biochemical)


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  • 99
    Name:
    Deoxyribonuclease I
    Description:
    Chromatographically purified A lyophilized powder with glycine as a stabilizer
    Catalog Number:
    ls002004
    Price:
    33
    Size:
    5 mg
    Source:
    Bovine Pancreas
    Cas Number:
    9003.98.9
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    Structured Review

    Worthington Biochemical dnase i
    Constitutive binding to interphase chromatin of an NFAT5a mutant lacking the CTD. (A) Flow cytometry analysis of the cell cycle profile ( left histogram ) and proportion of interphase and mitotic cells ( dot plot ) in asynchronous HEK293 cultures. (B) HEK293 cells cultured in isotonic medium (310 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours were lysed and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. One set of samples was treated with <t>DNase</t> I during lysis, which caused the release of chromatin-associated proteins to the soluble fraction. NFAT5 and markers of soluble (pyruvate kinase) and chromatin-associated proteins (histone H3) were detected by Western blotting. (C) HEK293 cells expressing Myc-tagged full length NFAT5a (FL5), a DNA-binding mutant (FL5 DB1 ), or a construct comprising the amino-terminal region plus DNA-binding domain (ND5) (diagram in Figure 1B ) and its DNA-binding mutant (ND5 DB1 ) were cultured in isotonic medium (290 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours, lysed, and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. NFAT5 constructs were detected by Western blotting with an anti-Myc antibody. Results shown are representative of four independent experiments.
    Chromatographically purified A lyophilized powder with glycine as a stabilizer
    https://www.bioz.com/result/dnase i/product/Worthington Biochemical
    Average 99 stars, based on 977 article reviews
    Price from $9.99 to $1999.99
    dnase i - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "Exclusion of NFAT5 from Mitotic Chromatin Resets Its Nucleo-Cytoplasmic Distribution in Interphase"

    Article Title: Exclusion of NFAT5 from Mitotic Chromatin Resets Its Nucleo-Cytoplasmic Distribution in Interphase

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007036

    Constitutive binding to interphase chromatin of an NFAT5a mutant lacking the CTD. (A) Flow cytometry analysis of the cell cycle profile ( left histogram ) and proportion of interphase and mitotic cells ( dot plot ) in asynchronous HEK293 cultures. (B) HEK293 cells cultured in isotonic medium (310 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours were lysed and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. One set of samples was treated with DNase I during lysis, which caused the release of chromatin-associated proteins to the soluble fraction. NFAT5 and markers of soluble (pyruvate kinase) and chromatin-associated proteins (histone H3) were detected by Western blotting. (C) HEK293 cells expressing Myc-tagged full length NFAT5a (FL5), a DNA-binding mutant (FL5 DB1 ), or a construct comprising the amino-terminal region plus DNA-binding domain (ND5) (diagram in Figure 1B ) and its DNA-binding mutant (ND5 DB1 ) were cultured in isotonic medium (290 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours, lysed, and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. NFAT5 constructs were detected by Western blotting with an anti-Myc antibody. Results shown are representative of four independent experiments.
    Figure Legend Snippet: Constitutive binding to interphase chromatin of an NFAT5a mutant lacking the CTD. (A) Flow cytometry analysis of the cell cycle profile ( left histogram ) and proportion of interphase and mitotic cells ( dot plot ) in asynchronous HEK293 cultures. (B) HEK293 cells cultured in isotonic medium (310 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours were lysed and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. One set of samples was treated with DNase I during lysis, which caused the release of chromatin-associated proteins to the soluble fraction. NFAT5 and markers of soluble (pyruvate kinase) and chromatin-associated proteins (histone H3) were detected by Western blotting. (C) HEK293 cells expressing Myc-tagged full length NFAT5a (FL5), a DNA-binding mutant (FL5 DB1 ), or a construct comprising the amino-terminal region plus DNA-binding domain (ND5) (diagram in Figure 1B ) and its DNA-binding mutant (ND5 DB1 ) were cultured in isotonic medium (290 mOsm/kg) or exposed to hypertonic conditions (470 mOsm/kg) during 4 hours, lysed, and proteins were fractionated into soluble (S) or chromatin (Ch) fractions. NFAT5 constructs were detected by Western blotting with an anti-Myc antibody. Results shown are representative of four independent experiments.

    Techniques Used: Binding Assay, Mutagenesis, Flow Cytometry, Cytometry, Cell Culture, Lysis, Western Blot, Expressing, Construct

    Related Articles

    Irradiation:

    Article Title: Characterization of the survival motor neuron (SMN) promoter provides evidence for complex combinatorial regulation in undifferentiated and differentiated P19 cells
    Article Snippet: .. Specifically, we used DMS, UVC irradiation and DNase I as DNA-modifying agents to map single-strand DNA breaks, comparing in vitro (naked DNA) and in vivo (living cells) footprints. .. In vivo footprints corresponding to nt −46 to +125 encompassing the minimal core promoter are presented in and summarized in .

    In Vivo:

    Article Title: Characterization of the survival motor neuron (SMN) promoter provides evidence for complex combinatorial regulation in undifferentiated and differentiated P19 cells
    Article Snippet: .. Specifically, we used DMS, UVC irradiation and DNase I as DNA-modifying agents to map single-strand DNA breaks, comparing in vitro (naked DNA) and in vivo (living cells) footprints. .. In vivo footprints corresponding to nt −46 to +125 encompassing the minimal core promoter are presented in and summarized in .

    In Vitro:

    Article Title: Characterization of the survival motor neuron (SMN) promoter provides evidence for complex combinatorial regulation in undifferentiated and differentiated P19 cells
    Article Snippet: .. Specifically, we used DMS, UVC irradiation and DNase I as DNA-modifying agents to map single-strand DNA breaks, comparing in vitro (naked DNA) and in vivo (living cells) footprints. .. In vivo footprints corresponding to nt −46 to +125 encompassing the minimal core promoter are presented in and summarized in .

    Produced:

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay
    Article Snippet: .. The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0. .. For evaluation of the quality of the assay, Z’ values were calculated using Equation 2: Z\u2019 =\u00a01\u00a0\u2212\u00a0(3SDC +\u00a03SDB )/(MC \u00a0\u2212\u00a0MB ) (2) where M = mean value; SD = standard deviation; C = control; and B = background.

    Concentration Assay:

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay
    Article Snippet: .. The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0. .. For evaluation of the quality of the assay, Z’ values were calculated using Equation 2: Z\u2019 =\u00a01\u00a0\u2212\u00a0(3SDC +\u00a03SDB )/(MC \u00a0\u2212\u00a0MB ) (2) where M = mean value; SD = standard deviation; C = control; and B = background.

    Incubation:

    Article Title: Identification of Viral Peptide Fragments for Vaccine Development
    Article Snippet: .. When the random digestion by DNase I is used for some genes, peculiarly a small amount of the full-length gene has been observed even after prolonged incubation. .. In this case, the digestion mixture should be passed through a column with an appropriate molecular weight cut-off to filter out the full-length gene, followed by a Microcon® YM-30 column to accomplish the buffer exchange.

    Article Title: Neutrophil Extracellular Traps Contain Calprotectin, a Cytosolic Protein Complex Involved in Host Defense against Candida albicans
    Article Snippet: .. As controls (i) NETs were mock-digested with nuclease-free RPMI and (ii) unstimulated neutrophils that did not release NETs were washed twice and incubated with RPMI containing 10 U/ml Dnase-1 for 20 min at 37°C. .. Four samples out of 4 wells were pooled, acetone precipitated, solubilized in 120 µl SDS loading buffer and boiled for 3 min. To account for potential protein loss due to proteolytic activity in the samples a complete purification procedure was performed in the presence of protease inhibitor cocktail (Sigma P1860; 1∶200) added to the wells 2 h after stimulation start as described above.

    other:

    Article Title: Transcriptional regulatory logic of the diurnal cycle in the mouse liver
    Article Snippet: Movie S2: Dynamics of DNase I, Pol II and H3K27ac at the Npas2 locus.

    Article Title: Identification of Viral Peptide Fragments for Vaccine Development
    Article Snippet: In the random digestion step, the use of DNase I in the presence of MnCl2 is critical as this protocol will generate DNA fragments of relatively uniform sizes, which facilitates the reassembly step ( 17 , also see Note ).

    Article Title: Neutrophil Extracellular Traps Contain Calprotectin, a Cytosolic Protein Complex Involved in Host Defense against Candida albicans
    Article Snippet: Supernatants were removed, NETs were washed twice with 1 ml RPMI and digested with 500 µl 10 U/ml DNase-1 each.

    Activity Assay:

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay
    Article Snippet: .. The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0. .. For evaluation of the quality of the assay, Z’ values were calculated using Equation 2: Z\u2019 =\u00a01\u00a0\u2212\u00a0(3SDC +\u00a03SDB )/(MC \u00a0\u2212\u00a0MB ) (2) where M = mean value; SD = standard deviation; C = control; and B = background.

    Staining:

    Article Title: Constitutive Nucleosome Depletion and Ordered Factor Assembly at the GRP78 Promoter Revealed by Single Molecule Footprinting
    Article Snippet: .. These were then digested at 37 °C for 15 min using various concentrations of DNase I (Worthington, San Francisco, California, United States) to obtain a suitable range of digestion of genomic DNA as revealed by EtBr staining. .. Digested genomic DNA was purified, redigested by RsaI, resolved on a 1.5% agarose gel, and Southern blotted.

    Recombinant:

    Article Title: Novel High-Throughput Deoxyribonuclease 1 Assay
    Article Snippet: .. The percentage of DNase I activity was calculated using Equation 1: DNase\u00a0I\u00a0activity (%) =\u00a0 (mean\u00a0velocity\u00a0of\u00a0a\u00a0compound/mean\u00a0velocity\u00a0of\u00a0DMSO)\u00a0\u00d7\u00a0100 (1) In similar assays, recombinant murine EndoG (produced in-house) was used at a concentration of 0.14 μM in 0.1 mM MgCl2 , 10 mM Tris-HCl, pH 7.4; and DNase II (Worthington, Lakewood, NJ) (3.32 nM) was tested in 100 mM sodium citrate buffer, pH 5.0. .. For evaluation of the quality of the assay, Z’ values were calculated using Equation 2: Z\u2019 =\u00a01\u00a0\u2212\u00a0(3SDC +\u00a03SDB )/(MC \u00a0\u2212\u00a0MB ) (2) where M = mean value; SD = standard deviation; C = control; and B = background.

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