protease k treatment  (Millipore)


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    Structured Review

    Millipore protease k treatment
    Protease K Treatment, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease k treatment/product/Millipore
    Average 98 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    protease k treatment - by Bioz Stars, 2020-03
    98/100 stars

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    Related Articles

    Raw Material:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals). .. Protocatechuate dehydrogenase (10 µg/mL; raw material purchased from Toyobo and further purified in-house to remove contaminating nucleases activity) ( ) was added to all mixtures immediately before injection into the imaging chamber.

    Amplification:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: In single-molecule protein-binding experiments, the fluorophores attached to the free end of DNA were usually removed by restriction digestion (an NdeI digestion site was introduced by the primer used for PCR amplification, and the removal was ∼95% efficient) after DNA mapping. .. All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Article Title: Reciprocal Role Of DNA Methylation And Sp1 Binding In Ki-67 Gene Transcription
    Article Snippet: After protease K treatment (Millipore), samples were extracted with phenol/chloroform and precipitated with ethanol. .. Recovered DNA was resuspended in TE buffer (Millipore) and amplified by PCR.

    DNA Synthesis:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: Paragraph title: DNA synthesis and ChIP. ... To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation.

    In Vitro:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: Paragraph title: In vitro single-molecule imaging and data analysis ... All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Protein Binding:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: In single-molecule protein-binding experiments, the fluorophores attached to the free end of DNA were usually removed by restriction digestion (an NdeI digestion site was introduced by the primer used for PCR amplification, and the removal was ∼95% efficient) after DNA mapping. .. All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Sonication:

    Article Title: Reciprocal Role Of DNA Methylation And Sp1 Binding In Ki-67 Gene Transcription
    Article Snippet: Chromatin Immunoprecipitation (ChIP) HK-2 and MKN45 cells were cross-linked by incubation in 1% formaldehyde-containing medium for 10 mins at 37°C, and sonicated to soluble chromatin. .. After protease K treatment (Millipore), samples were extracted with phenol/chloroform and precipitated with ethanol.

    Size-exclusion Chromatography:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: The slides were then cured for 2 min at 100°C before a second spin coating at 2000 rpm for 20 sec with a solution in toluene with the following: 0.5% polystyrene (Sigma, 182427), 0.005% azide-terminated polystyrene (Sigma, 699772), and M280 streptavidin beads (ThermoFisher, 11205D) (2 µL of beads for 200 µL of polystyrene solution prerinsed with methanol). .. All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Protease Inhibitor:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: The reaction was then mixed with 600 µl ChIP dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8, 16.7 mM NaCl, 1 mM PMSF, and protease inhibitor cocktail). .. To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation.

    Purification:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: .. All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals). .. Protocatechuate dehydrogenase (10 µg/mL; raw material purchased from Toyobo and further purified in-house to remove contaminating nucleases activity) ( ) was added to all mixtures immediately before injection into the imaging chamber.

    Real-time Polymerase Chain Reaction:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation. .. Sµ and Cµ sequences were quantified by quantitative PCR relative to input values.

    Incubation:

    Article Title: Reciprocal Role Of DNA Methylation And Sp1 Binding In Ki-67 Gene Transcription
    Article Snippet: Chromatin Immunoprecipitation (ChIP) HK-2 and MKN45 cells were cross-linked by incubation in 1% formaldehyde-containing medium for 10 mins at 37°C, and sonicated to soluble chromatin. .. After protease K treatment (Millipore), samples were extracted with phenol/chloroform and precipitated with ethanol.

    Article Title: Detection of Staphylococcus aureus with a fluorescence in situ hybridization that does not require lysostaphin
    Article Snippet: .. The proteinase K treatment replaced the 30 min lysozyme step with 10 min incubation in 1 mg/ml proteinase K (P4850, Sigma) at 40°C, a methanol rinse for inactivation, and 10 min incubation with 1 mg/ml lysozyme at 40°C. .. If not listed, other FISH steps were the same as the lysozyme‐only treatment.

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: .. To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation. .. Sµ and Cµ sequences were quantified by quantitative PCR relative to input values.

    Activity Assay:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals). .. Protocatechuate dehydrogenase (10 µg/mL; raw material purchased from Toyobo and further purified in-house to remove contaminating nucleases activity) ( ) was added to all mixtures immediately before injection into the imaging chamber.

    Imaging:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: Paragraph title: In vitro single-molecule imaging and data analysis ... All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Ethanol Precipitation:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: .. To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation. .. Sµ and Cµ sequences were quantified by quantitative PCR relative to input values.

    Polymerase Chain Reaction:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: In single-molecule protein-binding experiments, the fluorophores attached to the free end of DNA were usually removed by restriction digestion (an NdeI digestion site was introduced by the primer used for PCR amplification, and the removal was ∼95% efficient) after DNA mapping. .. All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals).

    Article Title: Reciprocal Role Of DNA Methylation And Sp1 Binding In Ki-67 Gene Transcription
    Article Snippet: After protease K treatment (Millipore), samples were extracted with phenol/chloroform and precipitated with ethanol. .. Recovered DNA was resuspended in TE buffer (Millipore) and amplified by PCR.

    Lysis:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: The cells were washed twice with 5 ml of cold phosphate-buffered saline supplemented with protease inhibitor cocktail tablet (Roche), lysed in 400 µl SDS lysis buffer (1% SDS, 10 mM EDTA, and 50 mM Tris-HCl, pH 8), and incubated on ice for 10 min. .. To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation.

    Injection:

    Article Title: Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis
    Article Snippet: All protein mixtures were made with a buffer containing 4.5% glycerol, 11.3 mM HEPES (pH 7.9), 5.6 mM MgCl2 , 45 mM KCl, 45 µM EDTA, 3.6 ng/µL yeast tRNA (Sigma) (additionally purified with protease K treatment and phenol/chloroform extraction), 0.1 ng/µL poly(dG:dC), 0.9 mM Trolox (Sigma), 2.3 mM protocatechuic acid (Sigma), 0.09% Tween 20 (EMD Chemicals), and 0.0045% NP40 (EMD Chemicals). .. Protocatechuate dehydrogenase (10 µg/mL; raw material purchased from Toyobo and further purified in-house to remove contaminating nucleases activity) ( ) was added to all mixtures immediately before injection into the imaging chamber.

    Binding Assay:

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: .. To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation. .. Sµ and Cµ sequences were quantified by quantitative PCR relative to input values.

    Fluorescence In Situ Hybridization:

    Article Title: Detection of Staphylococcus aureus with a fluorescence in situ hybridization that does not require lysostaphin
    Article Snippet: Fixatives and permeabilizers were selected on the basis of previous reports and the signal intensity, cells stained with FISH, cell loss after FISH, time taken for the assay, and costs were compared (Table ), for different permeabilization treatments. .. The proteinase K treatment replaced the 30 min lysozyme step with 10 min incubation in 1 mg/ml proteinase K (P4850, Sigma) at 40°C, a methanol rinse for inactivation, and 10 min incubation with 1 mg/ml lysozyme at 40°C.

    Chromatin Immunoprecipitation:

    Article Title: Reciprocal Role Of DNA Methylation And Sp1 Binding In Ki-67 Gene Transcription
    Article Snippet: Paragraph title: Chromatin Immunoprecipitation (ChIP) ... After protease K treatment (Millipore), samples were extracted with phenol/chloroform and precipitated with ethanol.

    Article Title: XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes
    Article Snippet: Paragraph title: DNA synthesis and ChIP. ... To get rid of nonspecific IgG binding proteins, the remaining cell suspension was incubated with 36 µl Dynabeads Protein G (Invitrogen) and 5 µg of normal mouse IgG antibody (12–371; Millipore) at 4°C for 3 h. The suspension was then transferred into fresh 1.5-ml tubes and incubated with 36 µl Dynabeads Protein G and 5 µg rabbit anti-XRCC1 antibody (ab9147; Abcam) at 4°C for 16 h. DNA was washed and eluted, and the cross-links were reversed by heating the Dynabeads at 65°C for 5 h followed by protease K treatment at 45°C for 2 h. DNA was recovered by phenol:chloroform:isoamyl alcohol 25:24:1 (Sigma-Aldrich) extraction and ethanol precipitation.

    Staining:

    Article Title: Detection of Staphylococcus aureus with a fluorescence in situ hybridization that does not require lysostaphin
    Article Snippet: Fixatives and permeabilizers were selected on the basis of previous reports and the signal intensity, cells stained with FISH, cell loss after FISH, time taken for the assay, and costs were compared (Table ), for different permeabilization treatments. .. The proteinase K treatment replaced the 30 min lysozyme step with 10 min incubation in 1 mg/ml proteinase K (P4850, Sigma) at 40°C, a methanol rinse for inactivation, and 10 min incubation with 1 mg/ml lysozyme at 40°C.

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  • 99
    Millipore proteinase k
    Proteinase K, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 610 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/proteinase k/product/Millipore
    Average 99 stars, based on 610 article reviews
    Price from $9.99 to $1999.99
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    https://www.bioz.com/result/protease k treatment/product/Millipore
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    Price from $9.99 to $1999.99
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    Millipore caspase 8 inhibitor
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