dtt  (New England Biolabs)


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  • 86
    Name:
    DNase I RNase free
    Description:
    DNase I RNase free 5 000 units
    Catalog Number:
    M0303L
    Price:
    282
    Category:
    Deoxyribonucleases DNase
    Size:
    5 000 units
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    Structured Review

    New England Biolabs dtt
    DNase I RNase free
    DNase I RNase free 5 000 units
    https://www.bioz.com/result/dtt/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dtt - by Bioz Stars, 2021-05
    86/100 stars

    Images

    Related Articles

    Footprinting:

    Article Title: The crystal structure of the TetR family transcriptional repressor SimR bound to DNA and the role of a flexible N-terminal extension in minor groove binding
    Article Snippet: After incubation at 22°C for 10 min, the binding reaction mixtures were loaded on 5% (w/v) native polyacrylamide gels and run in TBE buffer at 100 V for 45 min. EMSA data were collected and analysed on a PhosphoImager (FujiFilm) using Multi Gauge image analysis software (FujiFilm). .. DNase I footprinting Templates for DNase I footprinting were amplified by PCR using one unlabelled primer and one primer 5′-end labelled using [γ32 -P] ATP and T4 polynucleotide kinase (New England Biolabs). ..

    Amplification:

    Article Title: The crystal structure of the TetR family transcriptional repressor SimR bound to DNA and the role of a flexible N-terminal extension in minor groove binding
    Article Snippet: After incubation at 22°C for 10 min, the binding reaction mixtures were loaded on 5% (w/v) native polyacrylamide gels and run in TBE buffer at 100 V for 45 min. EMSA data were collected and analysed on a PhosphoImager (FujiFilm) using Multi Gauge image analysis software (FujiFilm). .. DNase I footprinting Templates for DNase I footprinting were amplified by PCR using one unlabelled primer and one primer 5′-end labelled using [γ32 -P] ATP and T4 polynucleotide kinase (New England Biolabs). ..

    Polymerase Chain Reaction:

    Article Title: The crystal structure of the TetR family transcriptional repressor SimR bound to DNA and the role of a flexible N-terminal extension in minor groove binding
    Article Snippet: After incubation at 22°C for 10 min, the binding reaction mixtures were loaded on 5% (w/v) native polyacrylamide gels and run in TBE buffer at 100 V for 45 min. EMSA data were collected and analysed on a PhosphoImager (FujiFilm) using Multi Gauge image analysis software (FujiFilm). .. DNase I footprinting Templates for DNase I footprinting were amplified by PCR using one unlabelled primer and one primer 5′-end labelled using [γ32 -P] ATP and T4 polynucleotide kinase (New England Biolabs). ..

    Lysis:

    Article Title: Detecting RNA-RNA interactions in E. coli using a modified CLASH method
    Article Snippet: Identification of ligated RNAs This study (Fig. ) employed in vivo crosslinking of RNA duplexes with the AMT molecule, which can, upon 365 nm UV irradiation, generate inter-strand adducts between juxtaposed uridine bases [ ]. .. Following cell lysis and RNA extraction, DNA residues were digested by DNase I, and single strand RNAs and free RNA overhangs adjacent to duplexes were digested by RNase T1. .. Then, the residual single strand RNAs were hybridized with 20 nt oligonucleotides and digested by RNase H three times.

    RNA Extraction:

    Article Title: Detecting RNA-RNA interactions in E. coli using a modified CLASH method
    Article Snippet: Identification of ligated RNAs This study (Fig. ) employed in vivo crosslinking of RNA duplexes with the AMT molecule, which can, upon 365 nm UV irradiation, generate inter-strand adducts between juxtaposed uridine bases [ ]. .. Following cell lysis and RNA extraction, DNA residues were digested by DNase I, and single strand RNAs and free RNA overhangs adjacent to duplexes were digested by RNase T1. .. Then, the residual single strand RNAs were hybridized with 20 nt oligonucleotides and digested by RNase H three times.

    Binding Assay:

    Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
    Article Snippet: Individual footprint reactions were initiated with a 22 μl binding reaction mixture that contained 12.5 mM HEPES (pH 7.8), 5 mM K-acetate (pH 8.0), 2.5 mM Mg-acetate, 1 mM CaCl2 , 12.5 μg/ml bovine serum albumin [ ], 0.3 mg/ml heparin, 200 nM of fluorescence-labeled DNA probe and various amounts of purified protein. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion. ..

    Incubation:

    Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
    Article Snippet: Individual footprint reactions were initiated with a 22 μl binding reaction mixture that contained 12.5 mM HEPES (pH 7.8), 5 mM K-acetate (pH 8.0), 2.5 mM Mg-acetate, 1 mM CaCl2 , 12.5 μg/ml bovine serum albumin [ ], 0.3 mg/ml heparin, 200 nM of fluorescence-labeled DNA probe and various amounts of purified protein. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion. ..

    Article Title: Intronic Cis-Regulatory Modules Mediate Tissue-Specific and Microbial Control of angptl4/fiaf Transcription
    Article Snippet: .. Nuclei were incubated with various concentrations of Dnase I (0–1.5 units, NEB) for 10 minutes at 37°C. .. Reactions were stopped by adding an equal volume of 2× Lysis Buffer (1% SDS, 200 mM NaCl, 10 mM EDTA, 20 mM Tris pH 7.5, 0.4 mg/ml proteinase K) and incubated overnight at 37°C.

    Synthesized:

    Article Title: Genome-wide characterization of methylguanosine-capped and polyadenylated small RNAs in the rice blast fungus Magnaporthe oryzae
    Article Snippet: The quantity of 5′-methylguanosine-capped RNA was measured by NanoDrop (Thermo Fisher) analysis and its integrity was determined with an Agilent 2100 Bioanalyzer. .. 3′-RACE analysis of CPA-sRNAs using 5′-capped RNA 5′-methylguanosine-capped RNA was treated with DNase I (NEB) to remove any contaminating genomic DNA. cDNA was synthesized in 20 µl reactions by adding the following reagents: 1 µg of 5′-methylguanosine-capped RNA, 50 picomole of 3′-oligo(dT) 20 VN primer, 5 mM of dNTPs, 1 U of RNaseOut (Invitrogen) and 5 U of Superscript III (Invitrogen). ..

    other:

    Article Title: Campylobacter-Induced Interleukin-8 Secretion in Polarized Human Intestinal Epithelial Cells Requires Campylobacter-Secreted Cytolethal Distending Toxin- and Toll-Like Receptor-Mediated Activation of NF-?B ▿
    Article Snippet: However, treatment of the basolateral conditioned supernatant with DNase I did not significantly change its ability to induce IL-8 secretion (Fig. ).

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  • 99
    New England Biolabs t4 pnk
    Purification and activity assays of PNKP and Polβ. ( A ) Purified Polβ-His 6 (17 ng) and PNKP-His 6 (127 ng) from E. coli were subjected to PAGE and stained with Coomassie blue. ( B ) Incorporation of [α- 32 P]-dCTP by Polβ using APE1-generated product. ddC, di-deoxynucleotide; P, product. ( C ) Schematic of the preparation of S (substrate) and subsequent enzymatic reactions for testing PNKP activity. ( D ) Efficiency of oligonucleotide labeling, annealing, and ligation leading to S indicated in ( C ). ( E ) Fpg (NEB, 1 U) completely digested S and the 3’ phosphate was completely removed by PNKP (12.7 ng and 127 ng, lanes 2 and 3), or by <t>T4</t> PNK (NEB, 0.1 U and 1 U, lanes 7 and 8). NEIL2 (272 ng) only partially digested S and its 3’P was resistant to the PNKP phosphatase (lanes 4 and 5).
    T4 Pnk, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 pnk/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 pnk - by Bioz Stars, 2021-05
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    99
    New England Biolabs sds sample buffer plus dtt
    Mutations in the Arsp-FB24 Arth_1007 intein fail to restore activity. Only unspliced MIP precursor (109 kDa) was observed with the wild type intein or with inteins mutated at essential intein residues that restored the consensus amino acid at position B∶10 (Asn 65 ) or possible F∶4 positions (Asp 315 or Gly 311 ). Simply Blue Safe stained <t>SDS-PAGE</t> of soluble lysates after in vivo expression at 37°C (0) or after incubation in vitro at 37°C at the indicated pH values in the presence (+) or absence (−) of 50 mM <t>DTT.</t> Western Blots with anti-P sera failed to detect spliced MP or cleavage products in all samples (data not shown). Molecular weight standards are in the left lane of each gel and selected sizes are listed in kDa (New England Biolabs 10 to 250 kDa ladder).
    Sds Sample Buffer Plus Dtt, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sds sample buffer plus dtt/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sds sample buffer plus dtt - by Bioz Stars, 2021-05
    99/100 stars
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    Image Search Results


    Purification and activity assays of PNKP and Polβ. ( A ) Purified Polβ-His 6 (17 ng) and PNKP-His 6 (127 ng) from E. coli were subjected to PAGE and stained with Coomassie blue. ( B ) Incorporation of [α- 32 P]-dCTP by Polβ using APE1-generated product. ddC, di-deoxynucleotide; P, product. ( C ) Schematic of the preparation of S (substrate) and subsequent enzymatic reactions for testing PNKP activity. ( D ) Efficiency of oligonucleotide labeling, annealing, and ligation leading to S indicated in ( C ). ( E ) Fpg (NEB, 1 U) completely digested S and the 3’ phosphate was completely removed by PNKP (12.7 ng and 127 ng, lanes 2 and 3), or by T4 PNK (NEB, 0.1 U and 1 U, lanes 7 and 8). NEIL2 (272 ng) only partially digested S and its 3’P was resistant to the PNKP phosphatase (lanes 4 and 5).

    Journal: eLife

    Article Title: Perturbation of base excision repair sensitizes breast cancer cells to APOBEC3 deaminase-mediated mutations

    doi: 10.7554/eLife.51605

    Figure Lengend Snippet: Purification and activity assays of PNKP and Polβ. ( A ) Purified Polβ-His 6 (17 ng) and PNKP-His 6 (127 ng) from E. coli were subjected to PAGE and stained with Coomassie blue. ( B ) Incorporation of [α- 32 P]-dCTP by Polβ using APE1-generated product. ddC, di-deoxynucleotide; P, product. ( C ) Schematic of the preparation of S (substrate) and subsequent enzymatic reactions for testing PNKP activity. ( D ) Efficiency of oligonucleotide labeling, annealing, and ligation leading to S indicated in ( C ). ( E ) Fpg (NEB, 1 U) completely digested S and the 3’ phosphate was completely removed by PNKP (12.7 ng and 127 ng, lanes 2 and 3), or by T4 PNK (NEB, 0.1 U and 1 U, lanes 7 and 8). NEIL2 (272 ng) only partially digested S and its 3’P was resistant to the PNKP phosphatase (lanes 4 and 5).

    Article Snippet: The products were purified by PCI extraction and ethanol precipitation and treated with PNKP (25 mM HEPES-KOH, pH 7.6, 50 mM NaCl, 0.5 mM EDTA, 0.5 mM DTT, 100 ng/ml BSA, 5% Glycerol) or T4 PNK (70 mM Tris-HCl, 10 mM MgCl2 , 5 mM DTT, pH 6.0) at 37°C for 30 min.

    Techniques: Purification, Activity Assay, Polyacrylamide Gel Electrophoresis, Staining, Generated, Oligonucleotide Labeling, Ligation

    Mutations in the Arsp-FB24 Arth_1007 intein fail to restore activity. Only unspliced MIP precursor (109 kDa) was observed with the wild type intein or with inteins mutated at essential intein residues that restored the consensus amino acid at position B∶10 (Asn 65 ) or possible F∶4 positions (Asp 315 or Gly 311 ). Simply Blue Safe stained SDS-PAGE of soluble lysates after in vivo expression at 37°C (0) or after incubation in vitro at 37°C at the indicated pH values in the presence (+) or absence (−) of 50 mM DTT. Western Blots with anti-P sera failed to detect spliced MP or cleavage products in all samples (data not shown). Molecular weight standards are in the left lane of each gel and selected sizes are listed in kDa (New England Biolabs 10 to 250 kDa ladder).

    Journal: PLoS ONE

    Article Title: The Arthrobacter Species FB24 Arth_1007 (DnaB) Intein Is a Pseudogene

    doi: 10.1371/journal.pone.0026361

    Figure Lengend Snippet: Mutations in the Arsp-FB24 Arth_1007 intein fail to restore activity. Only unspliced MIP precursor (109 kDa) was observed with the wild type intein or with inteins mutated at essential intein residues that restored the consensus amino acid at position B∶10 (Asn 65 ) or possible F∶4 positions (Asp 315 or Gly 311 ). Simply Blue Safe stained SDS-PAGE of soluble lysates after in vivo expression at 37°C (0) or after incubation in vitro at 37°C at the indicated pH values in the presence (+) or absence (−) of 50 mM DTT. Western Blots with anti-P sera failed to detect spliced MP or cleavage products in all samples (data not shown). Molecular weight standards are in the left lane of each gel and selected sizes are listed in kDa (New England Biolabs 10 to 250 kDa ladder).

    Article Snippet: Soluble lysates were boiled for 5 min in SDS Sample Buffer plus DTT (New England Biolabs), loaded onto 10–20% Tris Glycine polyacrylamide gels (Invitrogen, Carlsbad, CA) and either stained with Simply Blue Safe Stain (Invitrogen) or transferred to nitrocellulose membranes for Western Blot analysis with antisera against the Maltose Binding Protein (New England Biolabs), paramyosin or the His tag (Merck, Germany) as described previously , , .

    Techniques: Activity Assay, Staining, SDS Page, In Vivo, Expressing, Incubation, In Vitro, Western Blot, Molecular Weight

    In vitro activities of DsbG* mutants. Purified DsbC/DsbG/DsbG* samples were subjected to various in vitro assays. Negative control samples without protein, wt DsbC and wt DsbG control samples are shown as black bars ( lanes 1–3 ). Selected DsbG* mutants ( lanes 4–6 , dark gray bars ) and engineered DsbG* mutants ( lanes 7–10 , light gray bars ) are indicated. (A) Disulfide bond reductase activity. Protein samples were incubated with disulfide-bonded insulin and the rate of its reduction was followed at 650 nm in the presence of DTT. (B) Disulfide bond oxidase activity. Protein samples were incubated with fully reduced hirudin and the increase in DsbG fluorescence excitation at 295 nm was monitored over time. (C) Disulfide bond isomerase activity. Protein samples were incubated with scrambled hirudin and the folding of correctly folded hirudin was measured by HPLC. Horizontal gray bar indicates level of activity observed for wt DsbG. DTT, dithiothreitol; HPLC, high-performance liquid chromatography.

    Journal: Antioxidants & Redox Signaling

    Article Title: Converting a Sulfenic Acid Reductase into a Disulfide Bond Isomerase

    doi: 10.1089/ars.2014.6235

    Figure Lengend Snippet: In vitro activities of DsbG* mutants. Purified DsbC/DsbG/DsbG* samples were subjected to various in vitro assays. Negative control samples without protein, wt DsbC and wt DsbG control samples are shown as black bars ( lanes 1–3 ). Selected DsbG* mutants ( lanes 4–6 , dark gray bars ) and engineered DsbG* mutants ( lanes 7–10 , light gray bars ) are indicated. (A) Disulfide bond reductase activity. Protein samples were incubated with disulfide-bonded insulin and the rate of its reduction was followed at 650 nm in the presence of DTT. (B) Disulfide bond oxidase activity. Protein samples were incubated with fully reduced hirudin and the increase in DsbG fluorescence excitation at 295 nm was monitored over time. (C) Disulfide bond isomerase activity. Protein samples were incubated with scrambled hirudin and the folding of correctly folded hirudin was measured by HPLC. Horizontal gray bar indicates level of activity observed for wt DsbG. DTT, dithiothreitol; HPLC, high-performance liquid chromatography.

    Article Snippet: The pellet was resuspended in a 80 μl 4-acetamido-4′-maleimidylstilbene-2,2′-disulfonic acid (AMS) buffer (15 mM AMS [Cat. No. A485; Invitrogen], 1% sodium dodecyl sulfate (SDS), and 100 mM Tris HCl pH 8.0) using a shaker for 20 min at room temperature, followed by an incubation at 37°C for 40 min. To identify the oxidized and reduced states of DsbG, two controls were performed: (i) oxidized DsbG: the TCA precipitated pellet was resuspended in 80 μl of buffer without AMS (1% SDS, 100 mM Tris HCl pH 8.0) (ii) reduced DsbG: the TCA precipitated pellet was resuspended initially in a 200 μl DTT buffer (100 mM DTT [Cat. No. B7705; NEB], 1% SDS, 100 mM Tris HCl pH 8.0) for 20 min at room temperature under agitation.

    Techniques: In Vitro, Purification, Negative Control, Activity Assay, Incubation, Fluorescence, High Performance Liquid Chromatography

    The MIP* branched intermediate decays when treated with thiols. The N341A mutant yielded large amounts of MIP*, which was purified by affinity chromatography on amylose resin and adjusted to either pH 5 or pH 9. MIP* was then incubated at room temperature for 14 h in the presence (+) or absence (−) of 50 m m DTT ( A ) or in the presence (+) or absence (−) of 50 m m Ala, Cys, Ser, or Thr ( B ). A Coomassie Blue-stained SDS-PAGE is shown. Abbreviations are as in the legend for Fig. 2 , with the addition of the use of the single letter amino acid codes and S for New England Biolabs Broad Range Protein Marker. C , proposed pathway for thiol-induced cleavage of the Block G BI. Steps 1 and 2 are reversible, with the equilibrium in step 2 favoring the Block G BI. Step 3, which includes Asn cyclization and the O–N acyl shift, is irreversible. In the N341A mutant, Block G BI can be isolated and unfolded with urea, making Step 2 irreversible. The observed decay of the Block G BI in response to thiols is due to elimination of the Block F BI by thiols and the equilibrium between the 2 BI. The Cys branch point in Block F (Cys 320 in the MP-Be DnaB intein) and the Thr +1 branch point in Block G are indicated below the rectangles as C and T , respectively.

    Journal: The Journal of Biological Chemistry

    Article Title: Splicing of the Mycobacteriophage Bethlehem DnaB Intein

    doi: 10.1074/jbc.M109.069567

    Figure Lengend Snippet: The MIP* branched intermediate decays when treated with thiols. The N341A mutant yielded large amounts of MIP*, which was purified by affinity chromatography on amylose resin and adjusted to either pH 5 or pH 9. MIP* was then incubated at room temperature for 14 h in the presence (+) or absence (−) of 50 m m DTT ( A ) or in the presence (+) or absence (−) of 50 m m Ala, Cys, Ser, or Thr ( B ). A Coomassie Blue-stained SDS-PAGE is shown. Abbreviations are as in the legend for Fig. 2 , with the addition of the use of the single letter amino acid codes and S for New England Biolabs Broad Range Protein Marker. C , proposed pathway for thiol-induced cleavage of the Block G BI. Steps 1 and 2 are reversible, with the equilibrium in step 2 favoring the Block G BI. Step 3, which includes Asn cyclization and the O–N acyl shift, is irreversible. In the N341A mutant, Block G BI can be isolated and unfolded with urea, making Step 2 irreversible. The observed decay of the Block G BI in response to thiols is due to elimination of the Block F BI by thiols and the equilibrium between the 2 BI. The Cys branch point in Block F (Cys 320 in the MP-Be DnaB intein) and the Thr +1 branch point in Block G are indicated below the rectangles as C and T , respectively.

    Article Snippet: Samples were boiled for 5 min in sample buffer plus DTT (New England Biolabs), loaded onto 4–20% SDS-polyacrylamide gels (Invitrogen) and either stained with Coomassie Blue or transferred to nitrocellulose for Western blot analysis with an anti-MBP or anti-paramyosin antibody ( ).

    Techniques: Mutagenesis, Purification, Affinity Chromatography, Incubation, Staining, SDS Page, Marker, Blocking Assay, Isolation