rnase mix  (Thermo Fisher)


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    Structured Review

    Thermo Fisher rnase mix
    Rnase Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase mix/product/Thermo Fisher
    Average 95 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    rnase mix - by Bioz Stars, 2020-07
    95/100 stars

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    Related Articles

    Clone Assay:

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells
    Article Snippet: .. Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence. .. Index 1 = CGTGAT; index 2 = ACATCG; index 3 = GCCTAA; IDT custom synthesis) DNA Clean & Concentrator-5 Kit (Zymo Research, cat. no. D4003) 1N NaOH 5M NaCl 5× SSC (0.75M NaCl, 0.075M sodium citrate, Denhardts solution [0.1% Ficoll, 0.1% polyvinylpyrrolidone, 0.l% BSA]) Hyb buffer (5× SSC + 0.05% Tween-20) E-Gel EX Agarose Gels, 2% (ThermoFisher Scientific, cat. no. G4020–02) Novex® TBE Gels, 10%, 12 well (ThermoFisher Scientific, cat. no. EC62752BOX) 6 × DNA loading buffer (ThermoFisher Scientific, cat. no. R0611) TrackIt™ 10 bp DNA Ladder (ThermoFisher Scientific, cat. no. 10488–019) SYBR® Gold Nucleic Acid Gel Stain (10,000 × Concentrate in DMSO, ThermoFisher Scientific, cat. no. S-11494) Costar Spin-X Centrifuge Tube Filters (Cole-Parmer, cat. no. WU-01937–38) Gel elution buffer (0.3 M sodium acetate pH 5.2, 0.1% SDS) Herracell CO2 incubator (ThermoFisher Scientific) Universal 320R centrifuge (Hettich) Microcentrifuge 5424R (Eppendorf) Thermomixer R (Eppendorf) IX70 inverted phase-contrast microscope (Olympus) VortexGenie2 (Scientific Industries) Sonic Dismembrator (Model 100, ThermoFisher Scientific) UV Crosslinker (VWR) Qubit Fluorometer (ThermoFisher Scientific) Magnetic stand (MPC-S, ThermoFisher Scientific) Agilent Bioanalyzer 2100 (Agilent Technologies) Veriti 96-Well Thermal Cycler (ThermoFisher Scientific) MiSeq and HiSeq 2500 Ultra-High-Throughput Sequencing Systems (Illumina)

    Immunoprecipitation:

    Article Title: Varicella-Zoster Virus IE4 Protein Interacts with SR Proteins and Exports mRNAs through the TAP/NXF1 Pathway
    Article Snippet: .. HeLa nuclear extracts, treated or not for 30 min at 37°C with RNases A/T1 mix (Fermentas), were incubated with antibody-coupled beads for 4 h at 4°C in SR immunoprecipitation buffer (50 mM Tris-HCl, pH 7.4, 100 mM KCl, 5 mM MgCl2 , 0.1% NP-40, 10% glycerol). ..

    Article Title: Varicella-Zoster Virus IE4 Protein Interacts with SR Proteins and Exports mRNAs through the TAP/NXF1 Pathway
    Article Snippet: .. For TAP/NXF1 and Aly/REF immunoprecipitation, MeWo total extracts, treated or not for 45 min at 37°C with RNases A/T1 mix (Fermentas), were precleared for 1 h at 4°C with normal rabbit or mouse IgG (SantaCruz). .. Extracts were then incubated for 2 h at 4°C with anti-IE4 , anti-TAP/NXF1 (Abcam), anti-Aly/REF (Abcam), anti-GFP (SantaCruz) or anti-Flag (Sigma) antibodies as indicated.

    Purification:

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells
    Article Snippet: .. Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence. .. Index 1 = CGTGAT; index 2 = ACATCG; index 3 = GCCTAA; IDT custom synthesis) DNA Clean & Concentrator-5 Kit (Zymo Research, cat. no. D4003) 1N NaOH 5M NaCl 5× SSC (0.75M NaCl, 0.075M sodium citrate, Denhardts solution [0.1% Ficoll, 0.1% polyvinylpyrrolidone, 0.l% BSA]) Hyb buffer (5× SSC + 0.05% Tween-20) E-Gel EX Agarose Gels, 2% (ThermoFisher Scientific, cat. no. G4020–02) Novex® TBE Gels, 10%, 12 well (ThermoFisher Scientific, cat. no. EC62752BOX) 6 × DNA loading buffer (ThermoFisher Scientific, cat. no. R0611) TrackIt™ 10 bp DNA Ladder (ThermoFisher Scientific, cat. no. 10488–019) SYBR® Gold Nucleic Acid Gel Stain (10,000 × Concentrate in DMSO, ThermoFisher Scientific, cat. no. S-11494) Costar Spin-X Centrifuge Tube Filters (Cole-Parmer, cat. no. WU-01937–38) Gel elution buffer (0.3 M sodium acetate pH 5.2, 0.1% SDS) Herracell CO2 incubator (ThermoFisher Scientific) Universal 320R centrifuge (Hettich) Microcentrifuge 5424R (Eppendorf) Thermomixer R (Eppendorf) IX70 inverted phase-contrast microscope (Olympus) VortexGenie2 (Scientific Industries) Sonic Dismembrator (Model 100, ThermoFisher Scientific) UV Crosslinker (VWR) Qubit Fluorometer (ThermoFisher Scientific) Magnetic stand (MPC-S, ThermoFisher Scientific) Agilent Bioanalyzer 2100 (Agilent Technologies) Veriti 96-Well Thermal Cycler (ThermoFisher Scientific) MiSeq and HiSeq 2500 Ultra-High-Throughput Sequencing Systems (Illumina)

    Polymerase Chain Reaction:

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells
    Article Snippet: .. Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence. .. Index 1 = CGTGAT; index 2 = ACATCG; index 3 = GCCTAA; IDT custom synthesis) DNA Clean & Concentrator-5 Kit (Zymo Research, cat. no. D4003) 1N NaOH 5M NaCl 5× SSC (0.75M NaCl, 0.075M sodium citrate, Denhardts solution [0.1% Ficoll, 0.1% polyvinylpyrrolidone, 0.l% BSA]) Hyb buffer (5× SSC + 0.05% Tween-20) E-Gel EX Agarose Gels, 2% (ThermoFisher Scientific, cat. no. G4020–02) Novex® TBE Gels, 10%, 12 well (ThermoFisher Scientific, cat. no. EC62752BOX) 6 × DNA loading buffer (ThermoFisher Scientific, cat. no. R0611) TrackIt™ 10 bp DNA Ladder (ThermoFisher Scientific, cat. no. 10488–019) SYBR® Gold Nucleic Acid Gel Stain (10,000 × Concentrate in DMSO, ThermoFisher Scientific, cat. no. S-11494) Costar Spin-X Centrifuge Tube Filters (Cole-Parmer, cat. no. WU-01937–38) Gel elution buffer (0.3 M sodium acetate pH 5.2, 0.1% SDS) Herracell CO2 incubator (ThermoFisher Scientific) Universal 320R centrifuge (Hettich) Microcentrifuge 5424R (Eppendorf) Thermomixer R (Eppendorf) IX70 inverted phase-contrast microscope (Olympus) VortexGenie2 (Scientific Industries) Sonic Dismembrator (Model 100, ThermoFisher Scientific) UV Crosslinker (VWR) Qubit Fluorometer (ThermoFisher Scientific) Magnetic stand (MPC-S, ThermoFisher Scientific) Agilent Bioanalyzer 2100 (Agilent Technologies) Veriti 96-Well Thermal Cycler (ThermoFisher Scientific) MiSeq and HiSeq 2500 Ultra-High-Throughput Sequencing Systems (Illumina)

    Incubation:

    Article Title: RNA Binding of Ebola Virus VP30 Is Essential for Activating Viral Transcription
    Article Snippet: .. For RNase treatment, the cleared supernatant was incubated with 10 μl RNase A/T1 mix (1,000 U/ml; Thermo Fisher Scientific). ..

    Article Title: Varicella-Zoster Virus IE4 Protein Interacts with SR Proteins and Exports mRNAs through the TAP/NXF1 Pathway
    Article Snippet: .. HeLa nuclear extracts, treated or not for 30 min at 37°C with RNases A/T1 mix (Fermentas), were incubated with antibody-coupled beads for 4 h at 4°C in SR immunoprecipitation buffer (50 mM Tris-HCl, pH 7.4, 100 mM KCl, 5 mM MgCl2 , 0.1% NP-40, 10% glycerol). ..

    Article Title: Quantitation of circulating satellite RNAs in pancreatic cancer patients
    Article Snippet: .. Subsequently, 12.5 U of RNase A/T1 mix in 150 μl of digestion buffer (Ambion) was added and incubated at 37°C for 30 minutes to degrade unhybridized single-stranded RNAs, followed by incubation with 225 μl of an RNase inactivation/precipitation solution (Ambion) and 150 μl of ethanol for 2 hours at –20°C. .. Protected double-stranded RNAs, including tandem repeat RNAs with complementary probe RNAs and outer spike-in RNAs, were precipitated by centrifugation at 15,000 g for 15 minutes and resuspended in 9 μl of nuclease-free water.

    Article Title: Genome-wide analysis of human global and transcription-coupled excision repair of UV damage at single-nucleotide resolution
    Article Snippet: .. DNA pellets were resuspended in 45 µL of water and then incubated with 5 µL of RNase A/T1 mixture (Thermo, EN0551) for 1 h at 37°C. .. After phenol-chloroform extraction, purification through G50 filtration columns (GE Healthcare), and ethanol precipitation, the DNA was then used for ligation.

    Sequencing:

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells
    Article Snippet: .. Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence. .. Index 1 = CGTGAT; index 2 = ACATCG; index 3 = GCCTAA; IDT custom synthesis) DNA Clean & Concentrator-5 Kit (Zymo Research, cat. no. D4003) 1N NaOH 5M NaCl 5× SSC (0.75M NaCl, 0.075M sodium citrate, Denhardts solution [0.1% Ficoll, 0.1% polyvinylpyrrolidone, 0.l% BSA]) Hyb buffer (5× SSC + 0.05% Tween-20) E-Gel EX Agarose Gels, 2% (ThermoFisher Scientific, cat. no. G4020–02) Novex® TBE Gels, 10%, 12 well (ThermoFisher Scientific, cat. no. EC62752BOX) 6 × DNA loading buffer (ThermoFisher Scientific, cat. no. R0611) TrackIt™ 10 bp DNA Ladder (ThermoFisher Scientific, cat. no. 10488–019) SYBR® Gold Nucleic Acid Gel Stain (10,000 × Concentrate in DMSO, ThermoFisher Scientific, cat. no. S-11494) Costar Spin-X Centrifuge Tube Filters (Cole-Parmer, cat. no. WU-01937–38) Gel elution buffer (0.3 M sodium acetate pH 5.2, 0.1% SDS) Herracell CO2 incubator (ThermoFisher Scientific) Universal 320R centrifuge (Hettich) Microcentrifuge 5424R (Eppendorf) Thermomixer R (Eppendorf) IX70 inverted phase-contrast microscope (Olympus) VortexGenie2 (Scientific Industries) Sonic Dismembrator (Model 100, ThermoFisher Scientific) UV Crosslinker (VWR) Qubit Fluorometer (ThermoFisher Scientific) Magnetic stand (MPC-S, ThermoFisher Scientific) Agilent Bioanalyzer 2100 (Agilent Technologies) Veriti 96-Well Thermal Cycler (ThermoFisher Scientific) MiSeq and HiSeq 2500 Ultra-High-Throughput Sequencing Systems (Illumina)

    Recombinant:

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells
    Article Snippet: .. Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence. .. Index 1 = CGTGAT; index 2 = ACATCG; index 3 = GCCTAA; IDT custom synthesis) DNA Clean & Concentrator-5 Kit (Zymo Research, cat. no. D4003) 1N NaOH 5M NaCl 5× SSC (0.75M NaCl, 0.075M sodium citrate, Denhardts solution [0.1% Ficoll, 0.1% polyvinylpyrrolidone, 0.l% BSA]) Hyb buffer (5× SSC + 0.05% Tween-20) E-Gel EX Agarose Gels, 2% (ThermoFisher Scientific, cat. no. G4020–02) Novex® TBE Gels, 10%, 12 well (ThermoFisher Scientific, cat. no. EC62752BOX) 6 × DNA loading buffer (ThermoFisher Scientific, cat. no. R0611) TrackIt™ 10 bp DNA Ladder (ThermoFisher Scientific, cat. no. 10488–019) SYBR® Gold Nucleic Acid Gel Stain (10,000 × Concentrate in DMSO, ThermoFisher Scientific, cat. no. S-11494) Costar Spin-X Centrifuge Tube Filters (Cole-Parmer, cat. no. WU-01937–38) Gel elution buffer (0.3 M sodium acetate pH 5.2, 0.1% SDS) Herracell CO2 incubator (ThermoFisher Scientific) Universal 320R centrifuge (Hettich) Microcentrifuge 5424R (Eppendorf) Thermomixer R (Eppendorf) IX70 inverted phase-contrast microscope (Olympus) VortexGenie2 (Scientific Industries) Sonic Dismembrator (Model 100, ThermoFisher Scientific) UV Crosslinker (VWR) Qubit Fluorometer (ThermoFisher Scientific) Magnetic stand (MPC-S, ThermoFisher Scientific) Agilent Bioanalyzer 2100 (Agilent Technologies) Veriti 96-Well Thermal Cycler (ThermoFisher Scientific) MiSeq and HiSeq 2500 Ultra-High-Throughput Sequencing Systems (Illumina)

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  • 95
    Thermo Fisher rnase a t1 mix
    Optimization of ORF57 immunoprecipitation and RNase digestion. (A) ORF57-RNA complexes were immunoprecipitated by a rabbit anti-ORF57 antibody from cells extract of BCBL-1 cells treated with valproic acid for 24 h (input), normal rabbit IgG served as a negative control. The levels of ORF57 in input (1%) and immunoprecipitates (3%) in CLIP experiment were detected by Western blot with mouse anti-ORF57 antibody detecting both full-length ORF57 protein (upper band) and its caspase-cleavage product (lower band). (B) To determine an optimal RNase digestion condition, the immunoprecipitated ORF57 complexes were incubated with various amounts of RNase <t>A/T1</t> mix for 5 sec at room temperature and followed by proteinase K treatment and RNA extraction. The RNase A/T1 digestion efficiency was checked by RT-PCR for the remaining KSHV PAN RNA, a known ORF57 target.
    Rnase A T1 Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase a t1 mix/product/Thermo Fisher
    Average 95 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    rnase a t1 mix - by Bioz Stars, 2020-07
    95/100 stars
      Buy from Supplier

    91
    Thermo Fisher rnase cocktail enzyme mix
    Protein aggregation caused by enzymatic degradation of RNA, and effect of pre-hydrolysed RNA or <t>RNase</t> inhibition. a SDS-PAGE analysis of soluble (Supernatant) and aggregated (Pellet) proteins after treatment of mouse brain tissue lysate with increasing amounts of a mixture of RNase A and <t>T1</t> (A/T1) or vehicle (Ve-). b Agarose gel electrophoresis analysis of RNA isolated from RNase-treated human neuronal cell lysate. c - d Insoluble proteins collected by centrifugation after co-treatment of human neuronal cell lysate with RNase A and an RNase A inhibitor (RNasin, c ), or following the addition of RNA pre-hydrolysed by RNase A or alkaline hydrolysis (NaOH, d ).
    Rnase Cocktail Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase cocktail enzyme mix/product/Thermo Fisher
    Average 91 stars, based on 87 article reviews
    Price from $9.99 to $1999.99
    rnase cocktail enzyme mix - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    Image Search Results


    Optimization of ORF57 immunoprecipitation and RNase digestion. (A) ORF57-RNA complexes were immunoprecipitated by a rabbit anti-ORF57 antibody from cells extract of BCBL-1 cells treated with valproic acid for 24 h (input), normal rabbit IgG served as a negative control. The levels of ORF57 in input (1%) and immunoprecipitates (3%) in CLIP experiment were detected by Western blot with mouse anti-ORF57 antibody detecting both full-length ORF57 protein (upper band) and its caspase-cleavage product (lower band). (B) To determine an optimal RNase digestion condition, the immunoprecipitated ORF57 complexes were incubated with various amounts of RNase A/T1 mix for 5 sec at room temperature and followed by proteinase K treatment and RNA extraction. The RNase A/T1 digestion efficiency was checked by RT-PCR for the remaining KSHV PAN RNA, a known ORF57 target.

    Journal: Current protocols in microbiology

    Article Title: CLIP-seq to identify KSHV ORF57-binding RNA in host B cells

    doi: 10.1002/cpmc.3

    Figure Lengend Snippet: Optimization of ORF57 immunoprecipitation and RNase digestion. (A) ORF57-RNA complexes were immunoprecipitated by a rabbit anti-ORF57 antibody from cells extract of BCBL-1 cells treated with valproic acid for 24 h (input), normal rabbit IgG served as a negative control. The levels of ORF57 in input (1%) and immunoprecipitates (3%) in CLIP experiment were detected by Western blot with mouse anti-ORF57 antibody detecting both full-length ORF57 protein (upper band) and its caspase-cleavage product (lower band). (B) To determine an optimal RNase digestion condition, the immunoprecipitated ORF57 complexes were incubated with various amounts of RNase A/T1 mix for 5 sec at room temperature and followed by proteinase K treatment and RNA extraction. The RNase A/T1 digestion efficiency was checked by RT-PCR for the remaining KSHV PAN RNA, a known ORF57 target.

    Article Snippet: Protein A beads (EMD Millipore, cat. no. 16–125) RNase A/T1 mix (2 mg/ml of RNase A and 5000 U/ml of RNase T1, ThermoFisher Scientific, cat. no. EN0551) Recombinant Shrimp Alkaline Phosphatase (rSAP, 1000U/ml, New England Biolabs, cat. no. M0371S) Proteinase K (600 mAU/ml, EMD Millipore, cat. no. 71049) Proteinase K buffer (1× IP buffer supplemented with 1% SDS) Phase Lock Gel Light, 1.5 ml tubes (VWR, cat. no. 10052–164) 3M sodium acetate, pH 5.2 (Quality Biological, cat. no. 351-035-721EA) 70–75% (v/v) ethanol 100% (v/v) ethanol Agilent RNA 6000 Pico Kit (Agilent Technologies, cat. no. 5067–1513) Universal miRNA Cloning Linker (New England BioLabs, cat. no. S1315S) 50% PEG8000 (New England BioLabs, from the T4 RNA Ligase II kit) RNaseOUT (ThermoFisher Scientific, cat. no. 10777–019) T4 RNA Ligase 2, truncated KQ (200,000 units/ml, New England Biolabs, cat. no. M0373L) Agencourt RNAClean beads (Beckman Coulter, cat. no. A29168) Agencourt AMPure XP - PCR Purification beads (Beckman Coulter, cat. no. ) dNTP Mix, 10 mM each (Bioline, cat. no. BIO-39044) SuperScript III First-Strand Synthesis System (ThermoFisher Scientific, cat. no. 18080–051) CircLigase ssDNA ligase (Epicentre, cat. no. CL4111K) Phusion DNA polymerase kit (New England BioLabs, cat. no. M0530S) Reverse transcription primer (IDT custom synthesis) [5’-(Phos)-AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGC-(SpC18)-CACTCA-(SpC18)-TTCAGACGTGTGCTCTTCCGATCTATTGATGGTGCCTACAG-3’] riboPCR_F primer (5’-AATGATACGGCGACCACCGAGATCTACAC-3’, IDT custom synthesis) Indexed primers (5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGA-CGTGTGCTCTTCCG-3’) (‘NNNNNN’ denotes the index, of which each index has a unique sequence.

    Techniques: Immunoprecipitation, Negative Control, Cross-linking Immunoprecipitation, Western Blot, Incubation, Size-exclusion Chromatography, RNA Extraction, Reverse Transcription Polymerase Chain Reaction

    Protein aggregation in cell lysate supplemented with structural oligonucleotides and treated with RNase. Amount of protein aggregation observed after treating cell lysate from Jurkat T-cells with various oligonucleotides and RNase A/T1 (A/T1) and, if indicated, together with RNase H (A/T1 + H). Data are expressed as a percentage of the aggregation observed in samples treated with RANse A/T1 only and represent the mean ± s.d of three independent experiments. **= p

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein aggregation in cell lysate supplemented with structural oligonucleotides and treated with RNase. Amount of protein aggregation observed after treating cell lysate from Jurkat T-cells with various oligonucleotides and RNase A/T1 (A/T1) and, if indicated, together with RNase H (A/T1 + H). Data are expressed as a percentage of the aggregation observed in samples treated with RANse A/T1 only and represent the mean ± s.d of three independent experiments. **= p

    Article Snippet: Enzymes and reagents RNase T1 (AM2280), RNase V1 (AM2275), RNase A/T1 cocktail (EN0551), DNase I (2222), and Yeast t-RNA (15401-011) were from Thermo Fisher Scientific.

    Techniques:

    Protein renaturation: effects on enzymatic activity and effects of various polyanions. a Coomassie-stained SDS-PAGE gels of RNase-aggregated proteins from human neurons renatured with various polyanions. Input represents 1/10th of each sample, taken before the first centrifugation, and Pel 1 and Sup 1 are the pellet and the supernatant, respectively, recovered after the first centrifugation. Pel 2 represents the pellet obtained after treating the Sup 1 fraction with RNase A/T1 followed by centrifugation, please see experimental outline in Fig. 3a for a full description. b Renaturing of recombinant TDP-43 using increasing amounts of total RNA. Pel 1 represents aggregated protein after renaturing. Pel 2 is the aggregated protein after the soluble fraction (Sup 1) has been treated with RNase A/T. c Gel-electrophoresis analysis of proteins from human neurons renatured with vehicle or total RNA and then captured with ATP-agarose beads (ATP). Input represents 1/10th of each sample taken directly after renaturation, before the first centrifugation, and Blank represents the sample without any ATP-agarose beads present. d ATP-hydrolysing activity of RNase-aggregated proteins from Jurkat T-cells renatured in the presence (RNA) or absence (Ve-) of RNA. Data in c are expressed in arbitrary units (AU) and represent the mean ± s.d of three independent experiments. **= p

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein renaturation: effects on enzymatic activity and effects of various polyanions. a Coomassie-stained SDS-PAGE gels of RNase-aggregated proteins from human neurons renatured with various polyanions. Input represents 1/10th of each sample, taken before the first centrifugation, and Pel 1 and Sup 1 are the pellet and the supernatant, respectively, recovered after the first centrifugation. Pel 2 represents the pellet obtained after treating the Sup 1 fraction with RNase A/T1 followed by centrifugation, please see experimental outline in Fig. 3a for a full description. b Renaturing of recombinant TDP-43 using increasing amounts of total RNA. Pel 1 represents aggregated protein after renaturing. Pel 2 is the aggregated protein after the soluble fraction (Sup 1) has been treated with RNase A/T. c Gel-electrophoresis analysis of proteins from human neurons renatured with vehicle or total RNA and then captured with ATP-agarose beads (ATP). Input represents 1/10th of each sample taken directly after renaturation, before the first centrifugation, and Blank represents the sample without any ATP-agarose beads present. d ATP-hydrolysing activity of RNase-aggregated proteins from Jurkat T-cells renatured in the presence (RNA) or absence (Ve-) of RNA. Data in c are expressed in arbitrary units (AU) and represent the mean ± s.d of three independent experiments. **= p

    Article Snippet: Enzymes and reagents RNase T1 (AM2280), RNase V1 (AM2275), RNase A/T1 cocktail (EN0551), DNase I (2222), and Yeast t-RNA (15401-011) were from Thermo Fisher Scientific.

    Techniques: Activity Assay, Staining, SDS Page, Centrifugation, Recombinant, Nucleic Acid Electrophoresis

    Interaction of VP30 and VP35 is RNA dependent. (A) Co-IP analysis of FLAG-tagged VP30_wt with HA-tagged VP35 after RNase treatment. HEK-293 cells expressing VP30 FLAG and/or VP35 HA were lysed 48 h posttransfection. An aliquot was taken from cell lysates as control for protein expression levels (input). Two additional aliquots from each extract sample were processed in parallel by adding an RNase A/T1 mix to the first aliquot (+ RNase), but not to the second (− RNase), followed by precip itation of protein complexes for 2 h at 4°C using mouse anti-HA agarose (Sigma-Aldrich). Western blot analysis was performed using a mouse anti-FLAG M2 biotinylated antibody and Alexa Fluor 680-conjugated streptavidin (shown in red). HA-tagged VP35 was stained by rabbit anti-HA and IRDye 800-conjugated goat anti-rabbit (shown in green). Detection of proteins was obtained with the LiCor Odyssey Imaging System. The quantification of Western blot signals for VP30 coprecipitated by VP35 after or without RNase treatment (bar diagram on the right) was based on five independent experiments. VP30 FLAG amounts coimmunoprecipitated without RNase were set to 100%. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. (B) Co-IP analysis of FLAG-tagged VP35 with HA-tagged VP35 after or without RNase treatment. The procedure was identical to that described for panel A, except that anti-FLAG agarose (Sigma-Aldrich) was used for the co-IP, a mouse anti-HA biotinylated antibody and Alexa Fluor 680-conjugated streptavidin (shown in red) were used for Western blot detection of VP35 HA , and a rabbit anti-FLAG and IRDye 800-conjugated goat anti-rabbit (shown in green) antibodies were used for detection of VP35 FLAG . The quantification of Western blot signals for VP35 FLAG coprecipitated with VP35 HA after or without RNase treatment (bar diagram on the right) was based on five independent experiments. (C) Interaction of VP30 with VP35 is dependent on both proteins' ability to bind to RNA. Co-IP analysis of VP30 or VP35 with corresponding mutants that lack RNA binding activity. For VP35, we used a mutant (VP35mut HA ) lacking dsRNA binding activity owing to mutations R305A, K309A, and R312A. VP30_5LA FLAG was used as a negative RNA binding mutant to evaluate its interaction with VP35. Co-IP of protein complexes was performed as described for panel B. Single expression of the proteins used as control is shown in lanes 1 to 4. Lane 5, coexpression of VP30 FLAG with VP35 HA ; lane 6, coexpression of VP30 FLAG with VP35mut HA ; lane 7, coexpression of VP30_5LA FLAG with VP35 HA . Western blot signals of VP30 FLAG precipitated by VP35 HA versus VP35mut HA were quantified based on three independent experiments (upper bar diagram on the right); Western blot signals of VP30 FLAG or VP30_5LA FLAG precipitated by VP35 HA were quantified based on four independent experiments (lower bar diagram on the right). Amounts of precipitated wild-type proteins were set to 100%. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. Mock lanes, untransfected cells.

    Journal: Journal of Virology

    Article Title: RNA Binding of Ebola Virus VP30 Is Essential for Activating Viral Transcription

    doi: 10.1128/JVI.00271-16

    Figure Lengend Snippet: Interaction of VP30 and VP35 is RNA dependent. (A) Co-IP analysis of FLAG-tagged VP30_wt with HA-tagged VP35 after RNase treatment. HEK-293 cells expressing VP30 FLAG and/or VP35 HA were lysed 48 h posttransfection. An aliquot was taken from cell lysates as control for protein expression levels (input). Two additional aliquots from each extract sample were processed in parallel by adding an RNase A/T1 mix to the first aliquot (+ RNase), but not to the second (− RNase), followed by precip itation of protein complexes for 2 h at 4°C using mouse anti-HA agarose (Sigma-Aldrich). Western blot analysis was performed using a mouse anti-FLAG M2 biotinylated antibody and Alexa Fluor 680-conjugated streptavidin (shown in red). HA-tagged VP35 was stained by rabbit anti-HA and IRDye 800-conjugated goat anti-rabbit (shown in green). Detection of proteins was obtained with the LiCor Odyssey Imaging System. The quantification of Western blot signals for VP30 coprecipitated by VP35 after or without RNase treatment (bar diagram on the right) was based on five independent experiments. VP30 FLAG amounts coimmunoprecipitated without RNase were set to 100%. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. (B) Co-IP analysis of FLAG-tagged VP35 with HA-tagged VP35 after or without RNase treatment. The procedure was identical to that described for panel A, except that anti-FLAG agarose (Sigma-Aldrich) was used for the co-IP, a mouse anti-HA biotinylated antibody and Alexa Fluor 680-conjugated streptavidin (shown in red) were used for Western blot detection of VP35 HA , and a rabbit anti-FLAG and IRDye 800-conjugated goat anti-rabbit (shown in green) antibodies were used for detection of VP35 FLAG . The quantification of Western blot signals for VP35 FLAG coprecipitated with VP35 HA after or without RNase treatment (bar diagram on the right) was based on five independent experiments. (C) Interaction of VP30 with VP35 is dependent on both proteins' ability to bind to RNA. Co-IP analysis of VP30 or VP35 with corresponding mutants that lack RNA binding activity. For VP35, we used a mutant (VP35mut HA ) lacking dsRNA binding activity owing to mutations R305A, K309A, and R312A. VP30_5LA FLAG was used as a negative RNA binding mutant to evaluate its interaction with VP35. Co-IP of protein complexes was performed as described for panel B. Single expression of the proteins used as control is shown in lanes 1 to 4. Lane 5, coexpression of VP30 FLAG with VP35 HA ; lane 6, coexpression of VP30 FLAG with VP35mut HA ; lane 7, coexpression of VP30_5LA FLAG with VP35 HA . Western blot signals of VP30 FLAG precipitated by VP35 HA versus VP35mut HA were quantified based on three independent experiments (upper bar diagram on the right); Western blot signals of VP30 FLAG or VP30_5LA FLAG precipitated by VP35 HA were quantified based on four independent experiments (lower bar diagram on the right). Amounts of precipitated wild-type proteins were set to 100%. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. Mock lanes, untransfected cells.

    Article Snippet: For RNase treatment, the cleared supernatant was incubated with 10 μl RNase A/T1 mix (1,000 U/ml; Thermo Fisher Scientific).

    Techniques: Co-Immunoprecipitation Assay, Expressing, Western Blot, Staining, Imaging, RNA Binding Assay, Activity Assay, Mutagenesis, Binding Assay

    Protein aggregation caused by enzymatic degradation of RNA, and effect of pre-hydrolysed RNA or RNase inhibition. a SDS-PAGE analysis of soluble (Supernatant) and aggregated (Pellet) proteins after treatment of mouse brain tissue lysate with increasing amounts of a mixture of RNase A and T1 (A/T1) or vehicle (Ve-). b Agarose gel electrophoresis analysis of RNA isolated from RNase-treated human neuronal cell lysate. c - d Insoluble proteins collected by centrifugation after co-treatment of human neuronal cell lysate with RNase A and an RNase A inhibitor (RNasin, c ), or following the addition of RNA pre-hydrolysed by RNase A or alkaline hydrolysis (NaOH, d ).

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein aggregation caused by enzymatic degradation of RNA, and effect of pre-hydrolysed RNA or RNase inhibition. a SDS-PAGE analysis of soluble (Supernatant) and aggregated (Pellet) proteins after treatment of mouse brain tissue lysate with increasing amounts of a mixture of RNase A and T1 (A/T1) or vehicle (Ve-). b Agarose gel electrophoresis analysis of RNA isolated from RNase-treated human neuronal cell lysate. c - d Insoluble proteins collected by centrifugation after co-treatment of human neuronal cell lysate with RNase A and an RNase A inhibitor (RNasin, c ), or following the addition of RNA pre-hydrolysed by RNase A or alkaline hydrolysis (NaOH, d ).

    Article Snippet: Thawed or fresh cells were lysed in 80 µl Lysis Buffer 1 as described above and treated with RNase A/T1 for one hour at 37°C.

    Techniques: Inhibition, SDS Page, Agarose Gel Electrophoresis, Isolation, Centrifugation

    Protein aggregation in cell lysate supplemented with structural oligonucleotides and treated with RNase. Amount of protein aggregation observed after treating cell lysate from Jurkat T-cells with various oligonucleotides and RNase A/T1 (A/T1) and, if indicated, together with RNase H (A/T1 + H). Data are expressed as a percentage of the aggregation observed in samples treated with RANse A/T1 only and represent the mean ± s.d of three independent experiments. **= p

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein aggregation in cell lysate supplemented with structural oligonucleotides and treated with RNase. Amount of protein aggregation observed after treating cell lysate from Jurkat T-cells with various oligonucleotides and RNase A/T1 (A/T1) and, if indicated, together with RNase H (A/T1 + H). Data are expressed as a percentage of the aggregation observed in samples treated with RANse A/T1 only and represent the mean ± s.d of three independent experiments. **= p

    Article Snippet: Thawed or fresh cells were lysed in 80 µl Lysis Buffer 1 as described above and treated with RNase A/T1 for one hour at 37°C.

    Techniques:

    Protein aggregation caused by enzymatic degradation of RNA in mouse brain tissue lysate. Western blot analysis of aggregated proteins, collected by centrifugation, after treatment of mouse brain lysate with a mixture of RNase A and RNase T1 (RNase A/T1), or vechicle (Ve-). Input represents the starting material, Sup the supernatant (soluble fraction), and Pellet the aggregated fraction after centrifugation.

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein aggregation caused by enzymatic degradation of RNA in mouse brain tissue lysate. Western blot analysis of aggregated proteins, collected by centrifugation, after treatment of mouse brain lysate with a mixture of RNase A and RNase T1 (RNase A/T1), or vechicle (Ve-). Input represents the starting material, Sup the supernatant (soluble fraction), and Pellet the aggregated fraction after centrifugation.

    Article Snippet: Thawed or fresh cells were lysed in 80 µl Lysis Buffer 1 as described above and treated with RNase A/T1 for one hour at 37°C.

    Techniques: Western Blot, Centrifugation

    Protein renaturation: effects on enzymatic activity and effects of various polyanions. a Coomassie-stained SDS-PAGE gels of RNase-aggregated proteins from human neurons renatured with various polyanions. Input represents 1/10th of each sample, taken before the first centrifugation, and Pel 1 and Sup 1 are the pellet and the supernatant, respectively, recovered after the first centrifugation. Pel 2 represents the pellet obtained after treating the Sup 1 fraction with RNase A/T1 followed by centrifugation, please see experimental outline in Fig. 3a for a full description. b Renaturing of recombinant TDP-43 using increasing amounts of total RNA. Pel 1 represents aggregated protein after renaturing. Pel 2 is the aggregated protein after the soluble fraction (Sup 1) has been treated with RNase A/T. c Gel-electrophoresis analysis of proteins from human neurons renatured with vehicle or total RNA and then captured with ATP-agarose beads (ATP). Input represents 1/10th of each sample taken directly after renaturation, before the first centrifugation, and Blank represents the sample without any ATP-agarose beads present. d ATP-hydrolysing activity of RNase-aggregated proteins from Jurkat T-cells renatured in the presence (RNA) or absence (Ve-) of RNA. Data in c are expressed in arbitrary units (AU) and represent the mean ± s.d of three independent experiments. **= p

    Journal: bioRxiv

    Article Title: Enzymatic degradation of RNA causes widespread protein aggregation in cell and tissue lysates

    doi: 10.1101/841577

    Figure Lengend Snippet: Protein renaturation: effects on enzymatic activity and effects of various polyanions. a Coomassie-stained SDS-PAGE gels of RNase-aggregated proteins from human neurons renatured with various polyanions. Input represents 1/10th of each sample, taken before the first centrifugation, and Pel 1 and Sup 1 are the pellet and the supernatant, respectively, recovered after the first centrifugation. Pel 2 represents the pellet obtained after treating the Sup 1 fraction with RNase A/T1 followed by centrifugation, please see experimental outline in Fig. 3a for a full description. b Renaturing of recombinant TDP-43 using increasing amounts of total RNA. Pel 1 represents aggregated protein after renaturing. Pel 2 is the aggregated protein after the soluble fraction (Sup 1) has been treated with RNase A/T. c Gel-electrophoresis analysis of proteins from human neurons renatured with vehicle or total RNA and then captured with ATP-agarose beads (ATP). Input represents 1/10th of each sample taken directly after renaturation, before the first centrifugation, and Blank represents the sample without any ATP-agarose beads present. d ATP-hydrolysing activity of RNase-aggregated proteins from Jurkat T-cells renatured in the presence (RNA) or absence (Ve-) of RNA. Data in c are expressed in arbitrary units (AU) and represent the mean ± s.d of three independent experiments. **= p

    Article Snippet: Thawed or fresh cells were lysed in 80 µl Lysis Buffer 1 as described above and treated with RNase A/T1 for one hour at 37°C.

    Techniques: Activity Assay, Staining, SDS Page, Centrifugation, Recombinant, Nucleic Acid Electrophoresis