Ribolock ribonuclease rnase inhibitor

Manufactured by Thermo Fisher Scientific

Sourced in Lithuania

RiboLock ribonuclease (RNase) inhibitor is a recombinant protein that binds and inhibits the activity of RNases. It provides protection against RNA degradation in various molecular biology applications.

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Lab products found in correlation

Deoxyribonuclease 1 dnase 1, by Thermo Fisher Scientific (5 mentions) Experion, by Bio-Rad (4 mentions) Rnalater, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Rnalater solution, by Thermo Fisher Scientific (1 mentions)

5 protocols using ribolock ribonuclease rnase inhibitor

RNA Isolation and Purification Protocol

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The ipsilateral subcortical structures were placed in RNAlater (Ambion, Austin, TX, USA) solution for 24 h at 4 °C and then stored at −70 °C. Total RNA was isolated using TRI Reagent (MRC, Cincinnati, OH, USA) and acid guanidinium thiocyanate-phenol-chloroform extraction [32 (link)]. The isolated RNA was treated with deoxyribonuclease I (DNase I) (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania) in the presence of RiboLock ribonuclease (RNase) inhibitor (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania), according to the manufacturer’s recommended protocol. Deproteinization was performed using a 1:1 phenol:chloroform mixture. The isolated RNA was precipitated with sodium acetate (3.0 M, pH 5.2) and ethanol. The RNA integrity was checked using capillary electrophoresis (Experion, BioRad, Hercules, CA, USA). RNA integrity number (RIN) was at least 9.0.

Filippenkov I.B., Stavchansky V.V., Denisova A.E., Valieva L.V., Remizova J.A., Mozgovoy I.V., Zaytceva E.I., Gubsky L.V., Limborska S.A, & Dergunova L.V. (2021). Genome-Wide RNA-Sequencing Reveals Massive Circular RNA Expression Changes of the Neurotransmission Genes in the Rat Brain after Ischemia–Reperfusion. Genes, 12(12), 1870.

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RNA Extraction from Subcortex Tissue

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Tissues were placed in RNAlater solution (Ambion, Austin, TX, USA) for 24 h at 0 °C and then stored at –70 °C. The total RNA from the subcortex, was isolated using TRI Reagent (MRC, Cincinnati, OH, USA) and acid guanidinium thiocyanate–phenol–chloroform extraction [51 (link)]. The isolated RNA was treated with deoxyribonuclease I (DNase I) (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania) in the presence of RiboLock ribonuclease (RNase) inhibitor (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania), according to the manufacturer’s recommended protocol. Deproteinization was performed using a 1:1 phenol:chloroform mixture. The isolated RNA was precipitated with sodium acetate (3.0 M, pH 5.2) and ethanol. The RNA integrity was checked using capillary electrophoresis (Experion, BioRad, Hercules, CA, USA). The RNA integrity number (RIN) was at least 9.0.

Sudarkina O.Y., Filippenkov I.B., Stavchansky V.V., Denisova A.E., Yuzhakov V.V., Sevan’kaeva L.E., Valieva L.V., Remizova J.A., Dmitrieva V.G., Gubsky L.V., Myasoedov N.F., Limborska S.A, & Dergunova L.V. (2021). Brain Protein Expression Profile Confirms the Protective Effect of the ACTH(4–7)PGP Peptide (Semax) in a Rat Model of Cerebral Ischemia–Reperfusion. International Journal of Molecular Sciences, 22(12), 6179.

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RNA Isolation from Subcortical Tissue

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Tissues were placed in RNAlater solution for 24 h at 0 °C and then stored at −70 °C. Total RNA from the subcortex, was isolated using TRIzol reagent (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) and acid guanidinium thiocyanate-phenol-chloroform extraction [39 (link)]. The isolated RNA was treated with deoxyribonuclease I (DNase I) (Thermo Fisher Scientific) in the presence of RiboLock ribonuclease (RNase) inhibitor (Thermo Fisher Scientific), according to the manufacturer’s recommended protocol. Deproteinization was performed using a 1:1 phenol:chloroform mixture. The isolated RNA was precipitated with sodium acetate (3.0 M, pH 5.2) and ethanol. RNA integrity was checked using capillary electrophoresis (Experion, BioRad, Hercules, CA, USA). RNA integrity number (RIN) was at least 9.0.

Filippenkov I.B., Stavchansky V.V., Denisova A.E., Yuzhakov V.V., Sevan’kaeva L.E., Sudarkina O.Y., Dmitrieva V.G., Gubsky L.V., Myasoedov N.F., Limborska S.A, & Dergunova L.V. (2020). Novel Insights into the Protective Properties of ACTH(4-7)PGP (Semax) Peptide at the Transcriptome Level Following Cerebral Ischaemia–Reperfusion in Rats. Genes, 11(6), 681.

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Total RNA Extraction from Subcortex

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Total RNA from the subcortex, was isolated using TRIzol reagent (Invitrogen, Thermo Fisher Scientific) and acid guanidinium thiocyanate–phenol–chloroform extraction [49 (link)]. The isolated RNA was treated with deoxyribonuclease I (DNase I) (Thermo Fisher Scientific) in the presence of RiboLock ribonuclease (RNase) inhibitor (Thermo Fisher Scientific), according to the manufacturer’s recommended protocol. Deproteinization was performed using a 1:1 phenol:chloroform mixture. The isolated RNA was precipitated with sodium acetate (3.0 M, pH 5.2) and ethanol. RNA integrity was checked using capillary electrophoresis (Experion, BioRad, USA).

Dergunova L.V., Filippenkov I.B., Stavchansky V.V., Denisova A.E., Yuzhakov V.V., Mozerov S.A., Gubsky L.V, & Limborska S.A. (2018). Genome-wide transcriptome analysis using RNA-Seq reveals a large number of differentially expressed genes in a transient MCAO rat model. BMC Genomics, 19, 655.

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Hippocampal RNA Extraction and Analysis

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Rats were sacrificed 6 h after vehicle/peptide injection (4.5 h after ARS) and hippocampal samples were collected. Rats were decapitated and brains were quickly removed on an ice-cold plate. The hippocampi from left and right hemispheres were isolated and rapidly weighed.
Hippocampal tissues were placed in RNAlater (Ambion, Austin, TX, USA) solution for 24 h at 0 °C and then stored at –70 °C. Total RNA from the hippocampus was isolated using TRI Reagent (MRC, Cincinnati, OH, USA) and acid guanidinium thiocyanate–phenol–chloroform extraction [77 (link)]. The isolated RNA was treated with deoxyribonuclease I (DNase I) (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania) in the presence of RiboLock ribonuclease (RNase) inhibitor (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania), according to the manufacturer’s recommended protocol. Deproteinization was performed using a 1:1 phenol:chloroform mixture. The isolated RNA was precipitated with sodium acetate (3.0 M, pH 5.2) and ethanol. RNA integrity was checked using capillary electrophoresis (Experion, BioRad, Hercules, CA, USA). RNA integrity number (RIN) was at least 9.0.

Filippenkov I.B., Stavchansky V.V., Glazova N.Y., Sebentsova E.A., Remizova J.A., Valieva L.V., Levitskaya N.G., Myasoedov N.F., Limborska S.A, & Dergunova L.V. (2021). Antistress Action of Melanocortin Derivatives Associated with Correction of Gene Expression Patterns in the Hippocampus of Male Rats Following Acute Stress. International Journal of Molecular Sciences, 22(18), 10054.

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