rnase v1 enzyme  (Thermo Fisher)


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  • 99
    Name:
    RNase Cocktail Enzyme Mix
    Description:
    Ambion RNase Cocktail is a mixture of two highly purified ribonucleases RNase A 500 U⁄ml and RNase T1 20 000 U⁄ml and is free of DNase and nicking activities Use RNase Cocktail for all situations where it is desirable to degrade RNA i e plasmid minipreps and ribonuclease protection assays RNase Cocktail is supplied in 50 glycerol for maximum convenience Digestion of RNA with RNase A alone leaves fragments of RNA which are large enough to be visible on agarose gels and precipitate in ethanol RNase A cuts after C and U residues and RNase T1 cuts after G residues Consequently the mixture of both enzymes results in a reduction in RNA fragment size over the use of either alone Quality ControlRNase Cocktail Enzyme Mix is rigorously tested for contaminating nonspecific endonuclease exonuclease and protease activity Functionality is determined in a ribonuclease protection assay Unit Definitions RNase A One unit of RNase A is the amount required to give an increase in absorption at 286 nm of 0 0146 absorbance units per minute in a 1 mL volume Unit assay conditions 100 mM Tris acetate pH 6 5 1 mM EDTA and 1 mM cyclic 2 3 CMP RNase T1 100 Units of RNase T1 is the amount of enzyme that yields an increase in absorption at 260 nm of 0 01428 units per min at room temperature using 60 µg⁄mL yeast total RNA as a substrate We also now offer Ambion RNA Grade ribonucleases A V1 and T1 for use in RNA structure⁄function studies For more information see AM2274 RNase A 1 µg⁄ml AM2275 RNase V1 0 1 U⁄µl AM2283 RNase T1 1 U⁄µl
    Catalog Number:
    am2286
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Extraction|Maxiprep|Midiprep|Miniprep|Nuclease Protection Assays|Plasmid DNA Purification|Nucleic Acid Gel Electrophoresis & Blotting
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher rnase v1 enzyme
    AFM image of G4 dendriplexes prepared for 20 minutes attacked by <t>RNase</t> enzyme. (A) AFM image of hexagonal G4 dendriplexes prepared by mixing of G4 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 20 minutes at room temperature before loading onto the surface of freshly cleaved mica. AFM images of G4 dendriplexes after incubation with RNase <t>V1</t> enzyme for 1–28 minutes (B–F) shows separation of the adsorbed dendriplexes and degradation of the complexed siRNA molecules (dark spots) that increased with the increase in incubation time. Two dendriplexes (defined wit dotted circles) remained intact throughout the incubation time with RNase V1 enzyme suggesting the formation of individual compact particles. Scale bar in these images is 200 nm and the Z scale is 15 nm.
    Ambion RNase Cocktail is a mixture of two highly purified ribonucleases RNase A 500 U⁄ml and RNase T1 20 000 U⁄ml and is free of DNase and nicking activities Use RNase Cocktail for all situations where it is desirable to degrade RNA i e plasmid minipreps and ribonuclease protection assays RNase Cocktail is supplied in 50 glycerol for maximum convenience Digestion of RNA with RNase A alone leaves fragments of RNA which are large enough to be visible on agarose gels and precipitate in ethanol RNase A cuts after C and U residues and RNase T1 cuts after G residues Consequently the mixture of both enzymes results in a reduction in RNA fragment size over the use of either alone Quality ControlRNase Cocktail Enzyme Mix is rigorously tested for contaminating nonspecific endonuclease exonuclease and protease activity Functionality is determined in a ribonuclease protection assay Unit Definitions RNase A One unit of RNase A is the amount required to give an increase in absorption at 286 nm of 0 0146 absorbance units per minute in a 1 mL volume Unit assay conditions 100 mM Tris acetate pH 6 5 1 mM EDTA and 1 mM cyclic 2 3 CMP RNase T1 100 Units of RNase T1 is the amount of enzyme that yields an increase in absorption at 260 nm of 0 01428 units per min at room temperature using 60 µg⁄mL yeast total RNA as a substrate We also now offer Ambion RNA Grade ribonucleases A V1 and T1 for use in RNA structure⁄function studies For more information see AM2274 RNase A 1 µg⁄ml AM2275 RNase V1 0 1 U⁄µl AM2283 RNase T1 1 U⁄µl
    https://www.bioz.com/result/rnase v1 enzyme/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rnase v1 enzyme - by Bioz Stars, 2020-07
    99/100 stars

    Related Products / Commonly Used Together

    anti-gapdh sirna

    Images

    1) Product Images from "Visualizing the Attack of RNase Enzymes on Dendriplexes and Naked RNA Using Atomic Force Microscopy"

    Article Title: Visualizing the Attack of RNase Enzymes on Dendriplexes and Naked RNA Using Atomic Force Microscopy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0061710

    AFM image of G4 dendriplexes prepared for 20 minutes attacked by RNase enzyme. (A) AFM image of hexagonal G4 dendriplexes prepared by mixing of G4 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 20 minutes at room temperature before loading onto the surface of freshly cleaved mica. AFM images of G4 dendriplexes after incubation with RNase V1 enzyme for 1–28 minutes (B–F) shows separation of the adsorbed dendriplexes and degradation of the complexed siRNA molecules (dark spots) that increased with the increase in incubation time. Two dendriplexes (defined wit dotted circles) remained intact throughout the incubation time with RNase V1 enzyme suggesting the formation of individual compact particles. Scale bar in these images is 200 nm and the Z scale is 15 nm.
    Figure Legend Snippet: AFM image of G4 dendriplexes prepared for 20 minutes attacked by RNase enzyme. (A) AFM image of hexagonal G4 dendriplexes prepared by mixing of G4 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 20 minutes at room temperature before loading onto the surface of freshly cleaved mica. AFM images of G4 dendriplexes after incubation with RNase V1 enzyme for 1–28 minutes (B–F) shows separation of the adsorbed dendriplexes and degradation of the complexed siRNA molecules (dark spots) that increased with the increase in incubation time. Two dendriplexes (defined wit dotted circles) remained intact throughout the incubation time with RNase V1 enzyme suggesting the formation of individual compact particles. Scale bar in these images is 200 nm and the Z scale is 15 nm.

    Techniques Used: Incubation

    AFM image of G5 dendriplexes prepared for 20 minutes attacked by RNase enzyme. (A) AFM image of hexagonal G5 dendriplexes prepared by mixing of G5 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 20 minutes at room temperature before loading onto the surface of freshly cleaved mica. AFM images of G5 dendriplexes after incubation with RNase V1 enzyme for 1–60 minutes (B–F) shows separation of the adsorbed dendriplexes and degradation of the complexed siRNA molecules (dark spots) that increased with the increase in incubation time. Scale bar in these images is 200 nm and the Z scale is 17 nm.
    Figure Legend Snippet: AFM image of G5 dendriplexes prepared for 20 minutes attacked by RNase enzyme. (A) AFM image of hexagonal G5 dendriplexes prepared by mixing of G5 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 20 minutes at room temperature before loading onto the surface of freshly cleaved mica. AFM images of G5 dendriplexes after incubation with RNase V1 enzyme for 1–60 minutes (B–F) shows separation of the adsorbed dendriplexes and degradation of the complexed siRNA molecules (dark spots) that increased with the increase in incubation time. Scale bar in these images is 200 nm and the Z scale is 17 nm.

    Techniques Used: Incubation

    AFM image of free siRNAs before and after attack by RNase enzyme. (A) AFM image of free anti-GAPDH siRNA dissolved in 1 mM PBS containing 2 mM MgCl 2 after adding to the surface of freshly cleaved mica, which shows rod-, sphere-, and bead-like arrangements. (B) AFM image taken 1.5 minutes after adding RNase V1 enzyme, which shows rapid fragmentation of adsorbed siRNA molecules. (C) Time-lapse images showing a single siRNA molecule denoted by the white arrow ( t = 0 min), the attack of RNase V1 enzyme on free siRNA molecule ( t = 1.5 min), and complete siRNA degradation ( t = 3 min). The scale bar in images A and B is 100 nm and the Z scale is 9 nm. The scale bar in image C is 35 nm and Z scale is 7 nm.
    Figure Legend Snippet: AFM image of free siRNAs before and after attack by RNase enzyme. (A) AFM image of free anti-GAPDH siRNA dissolved in 1 mM PBS containing 2 mM MgCl 2 after adding to the surface of freshly cleaved mica, which shows rod-, sphere-, and bead-like arrangements. (B) AFM image taken 1.5 minutes after adding RNase V1 enzyme, which shows rapid fragmentation of adsorbed siRNA molecules. (C) Time-lapse images showing a single siRNA molecule denoted by the white arrow ( t = 0 min), the attack of RNase V1 enzyme on free siRNA molecule ( t = 1.5 min), and complete siRNA degradation ( t = 3 min). The scale bar in images A and B is 100 nm and the Z scale is 9 nm. The scale bar in image C is 35 nm and Z scale is 7 nm.

    Techniques Used:

    AFM image of G5 dendriplexes prepared for 24 hours attacked by RNase enzyme. (A) AFM image of G5 dendriplexes prepared by mixing of G5 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 24 hours at room temperature before loading onto the surface of freshly cleaved mica. G5 dendriplexes remain intact upon incubating with RNase V1 enzyme for 30 (B) and 60 minutes (C). Scale bar in these AFM images is 140 nm and the Z scale is 5 nm.
    Figure Legend Snippet: AFM image of G5 dendriplexes prepared for 24 hours attacked by RNase enzyme. (A) AFM image of G5 dendriplexes prepared by mixing of G5 dendrimers with 0.7 µg of anti-GAPDH siRNA at N/P ratio of 2/1 for 24 hours at room temperature before loading onto the surface of freshly cleaved mica. G5 dendriplexes remain intact upon incubating with RNase V1 enzyme for 30 (B) and 60 minutes (C). Scale bar in these AFM images is 140 nm and the Z scale is 5 nm.

    Techniques Used:

    Related Articles

    Magnetic Beads:

    Article Title: DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs
    Article Snippet: .. Co-Immunoprecipitation experiments FH-DND1-HEK293 cells from one confluent 15-cm plate were lysed in NP40 lysis buffer (50 mM HEPES, pH 7.5, 150 mM KCl, 2 mM MgCl2 , 1 mM NaF, 0.5% (v/v) NP40, 0.5 mM DTT, complete EDTA-free protease inhibitor cocktail (Roche)) and FH-DND1 immunoprecipitated with anti-FLAG-M2 magnetic beads (SIGMA-ALDRICH, cat #M8823) in the presence or absence of 5 U/μl RNase A and 200 U/μl RNase T1 (RNase A/T1 mix, Thermo-Fisher, cat #AM2286). .. Magnetic beads were washed twice with NP40 lysis buffer and the immunoprecipitate eluted with SDS-gel loading buffer.

    Protease Inhibitor:

    Article Title: DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs
    Article Snippet: .. Co-Immunoprecipitation experiments FH-DND1-HEK293 cells from one confluent 15-cm plate were lysed in NP40 lysis buffer (50 mM HEPES, pH 7.5, 150 mM KCl, 2 mM MgCl2 , 1 mM NaF, 0.5% (v/v) NP40, 0.5 mM DTT, complete EDTA-free protease inhibitor cocktail (Roche)) and FH-DND1 immunoprecipitated with anti-FLAG-M2 magnetic beads (SIGMA-ALDRICH, cat #M8823) in the presence or absence of 5 U/μl RNase A and 200 U/μl RNase T1 (RNase A/T1 mix, Thermo-Fisher, cat #AM2286). .. Magnetic beads were washed twice with NP40 lysis buffer and the immunoprecipitate eluted with SDS-gel loading buffer.

    Immunoprecipitation:

    Article Title: DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs
    Article Snippet: .. Co-Immunoprecipitation experiments FH-DND1-HEK293 cells from one confluent 15-cm plate were lysed in NP40 lysis buffer (50 mM HEPES, pH 7.5, 150 mM KCl, 2 mM MgCl2 , 1 mM NaF, 0.5% (v/v) NP40, 0.5 mM DTT, complete EDTA-free protease inhibitor cocktail (Roche)) and FH-DND1 immunoprecipitated with anti-FLAG-M2 magnetic beads (SIGMA-ALDRICH, cat #M8823) in the presence or absence of 5 U/μl RNase A and 200 U/μl RNase T1 (RNase A/T1 mix, Thermo-Fisher, cat #AM2286). .. Magnetic beads were washed twice with NP40 lysis buffer and the immunoprecipitate eluted with SDS-gel loading buffer.

    Incubation:

    Article Title: CryoEM structure of Saccharomyces cerevisiae U1 snRNP offers insight into alternative splicing
    Article Snippet: .. Beads were incubated with 100 µl lysis buffer containing 0.5 U RNase A and 20 U RNase T1 (Life Technologies, AM2286) or no RNase, at room temperature for 20 min. Beads were then washed with cell lysis buffer another six times. .. Half of beads were boiled with 2× SDS-sample buffer for SDS-PAGE and western blot (Supplementary Fig. ).

    Article Title: Pathogen-derived extracellular vesicles mediate virulence in the fatal human pathogen Cryptococcus gattii
    Article Snippet: .. To degrade RNA, 50 μl of EVsR265 (at protein concentration of around 6 mg ml−1 ) was incubated in the presence of 2.5 μl RNase cocktail of RNase A and T1 (RNase Cocktail™ Enzyme Mix, Thermo Fisher Scientific #AM2286) for 15 min at 37 °C. .. Degradation of nucleic acids was confirmed by isolation of nucleic acids from treated and untreated EVsR265 using Wizard Genomic DNA Purification Kit (Promega) without RNase A added and visualising them on 1% agarose gel.

    Article Title: Defining the RNA interactome by total RNA‐associated protein purification
    Article Snippet: .. Add 0.5 μl of RNaseA+T1 (Ambion AM2286) mix per tube and incubate the samples for 2 h in a SpeedVac at 45°C, followed by SpeedVac incubation at 65°C until the samples are fully dry. ..

    Article Title: The Human Nucleolar Protein FTSJ3 Associates with NIP7 and Functions in Pre-rRNA Processing
    Article Snippet: .. For RNA-dependent protein interaction experiments, RNase A + T1 cocktail (AM2286 – Ambion) was titrated down (2.5 U RNase A and 100 U RNase T1, 0.25 U RNase A and 10 U RNase T1, 0.025 U RNase A and 1 U RNase T1, 0.0025 U RNase A and 0.1 U RNase T1 and 0.00025 U RNase A and 0.01 U RNase T1), and the bead suspension incubated in a water bath at 37°C for 10 minutes, and subsequently cooled for 5 minutes on ice. .. The supernatant was saved and beads were washed 3 times with 1 ml of IsoWB and suspended in 50 µl of SDS-PAGE loading buffer .

    Protein Concentration:

    Article Title: Pathogen-derived extracellular vesicles mediate virulence in the fatal human pathogen Cryptococcus gattii
    Article Snippet: .. To degrade RNA, 50 μl of EVsR265 (at protein concentration of around 6 mg ml−1 ) was incubated in the presence of 2.5 μl RNase cocktail of RNase A and T1 (RNase Cocktail™ Enzyme Mix, Thermo Fisher Scientific #AM2286) for 15 min at 37 °C. .. Degradation of nucleic acids was confirmed by isolation of nucleic acids from treated and untreated EVsR265 using Wizard Genomic DNA Purification Kit (Promega) without RNase A added and visualising them on 1% agarose gel.

    Lysis:

    Article Title: CryoEM structure of Saccharomyces cerevisiae U1 snRNP offers insight into alternative splicing
    Article Snippet: .. Beads were incubated with 100 µl lysis buffer containing 0.5 U RNase A and 20 U RNase T1 (Life Technologies, AM2286) or no RNase, at room temperature for 20 min. Beads were then washed with cell lysis buffer another six times. .. Half of beads were boiled with 2× SDS-sample buffer for SDS-PAGE and western blot (Supplementary Fig. ).

    Article Title: DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs
    Article Snippet: .. Co-Immunoprecipitation experiments FH-DND1-HEK293 cells from one confluent 15-cm plate were lysed in NP40 lysis buffer (50 mM HEPES, pH 7.5, 150 mM KCl, 2 mM MgCl2 , 1 mM NaF, 0.5% (v/v) NP40, 0.5 mM DTT, complete EDTA-free protease inhibitor cocktail (Roche)) and FH-DND1 immunoprecipitated with anti-FLAG-M2 magnetic beads (SIGMA-ALDRICH, cat #M8823) in the presence or absence of 5 U/μl RNase A and 200 U/μl RNase T1 (RNase A/T1 mix, Thermo-Fisher, cat #AM2286). .. Magnetic beads were washed twice with NP40 lysis buffer and the immunoprecipitate eluted with SDS-gel loading buffer.

    Isolation:

    Article Title: Purification of Highly Active Alphavirus Replication Complexes Demonstrates Altered Fractionation of Multiple Cellular Membranes
    Article Snippet: .. Isolated RNA was treated with RNase cocktail enzyme mix (Ambion) containing RNase A and T1 (final concentrations of 2.5 and 100 U/ml, respectively) or RNase III (50 U/ml; NEBNext RNase III RNA fragmentation module) under high-salt conditions. ..

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    Thermo Fisher ultrapure bsa
    Ultrapure Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ultrapure bsa/product/Thermo Fisher
    Average 95 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    ultrapure bsa - by Bioz Stars, 2020-07
    95/100 stars
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