The RIBOPROTECT Hu RNase Inhibitor is a 50 kDa recombinant human placental protein expressed in Escherichia coli. It inhibits ribonuclease (RNase) activity of common eukaryotic enzymes such as RNase A, RNase B, RNase C, and protects RNA integrity from degradation. RIBOPROTECT Hu is intended for use in applications where the presence of RNases may cause a hazard to RNA quality and experiment results, e.g. in RNA isolation, cDNA synthesis, RT-PCR, in vitro transcription and translation, or RNase-free monoclonal antibody preparation. It is compatible with DNA Polymerases and AMV or M-MuLV Reverse Transcriptases.
Available Options:
| SKU | Unit | ||
|---|---|---|---|
 | RT35-020 | 2000 U ( 40U/μl ) | Ask for an offer |
| RT35-100 | 10 000 U ( 40U/μl ) | Ask for an offer |
The RIBOPROTECT Hu RNase Inhibitor is a 50 kDa recombinant human placental protein expressed in Escherichia coli. It inhibits ribonuclease (RNase) activity of common eukaryotic enzymes such as RNase A, RNase B, RNase C by non-covalent binding in a 1:1 ratio. RIBOPROTECT Hu is intended for use in applications where the presence of RNases may cause a hazard to RNA quality and experiment results, e.g. in RNA isolation, cDNA synthesis, RT-PCR, in vitro transcription and translation, or RNase-free monoclonal antibody preparation RIBOPROTECT Hu shows no activity towards RNase 1, RNase T1, RNase T2, S1 nuclease, and RNase H. It is compatible with DNA Polymerases and AMV or M-MuLV Reverse Transcriptases.
Features and advantages
- Completely inhibits RNase A, B and C activity
- No influence on the polymerase or reverse transcriptase activity
- Free of DNase and RNase activity
- Retains full stability at 37°C for at least 4 weeks
- Stable up to 58°C and at min. 0.5 – 1 mM DTT concentration ranges
- Active in diverse reaction conditions and in various buffers
- Active over a broad pH range (pH 5.5 – 9.0)
- Compatible with the TRANSCRIPTME Reverse Transcriptase (cat. no. RT32)
Applications
- RNA isolation and purification
- cDNA synthesis, RT-PCR, RT-qPCR
- in vitro transcription and translation
- RNase-free monoclonal antibody preparation
Unit definition
One unit is defined as the amount of enzyme required to inhibit the activity of 5 ng RNase A by 50%.
RIBOPROTECT Hu RNase Inhibitor (RT35) - Flyer
RIBOPROTECT Hu RNase Inhibitor (RT35) - Manual
RIBOPROTECT Hu RNase Inhibitor (RT35) - Flyer (Chinese)
RIBOPROTECT Hu RNase Inhibitor – Troubleshooting
Usage
- The optimal final concentration of the RIBOPROTECT Hu in a reaction depends on the level of RNase contamination, the incubation time and the compounds present in the reaction mixture. It falls within a range of 1 – 2 U/μl.
- For a standard reverse transcription reaction, use 1 μl (40 U) of the RIBOPROTECT Hu for a final sample volume of 20 μl.
- For the optimal RIBOPROTECT Hu activity, a final DTT concentration of 0.5 – 1 mM is essential.
- During assembly of a reaction, RIBOPROTECT Hu should be added before other components that are possible sources of RNase contamination.
- Using RIBOPROTECT Hu does not exclude RNase H treatment after amplification of the first strand cDNA.
Storage buffer
20 mM HEPES-KOH (pH 7.6); 50 mM KCl; 8 mM reducing agent; 50% (v/v) glycerol
Storage conditions
Store at -20°C in a freezer without a defrost cycle.
Quality control
The absence of Endonuclease, Exonuclease, RNase and latent RNase activities has been confirmed using the relevant procedures. The purity is >90% as judged by SDS-polyacrylamide gels.
RIBOPROTECT Hu RNase Inhibitor (RT35) - MSDS
RIBOPROTECT Hu RNase Inhibitor (RT35) - CoA
-
In Situ Sequencing: A High-Throughput, Multi-Targeted Gene Expression Profiling Technique for Cell Typing in Tissue Sections
Authors Markus M. Hilscher, Daniel Gyllborg, Chika Yokota, Mats Nilsson Products RIBOPROTECT Hu RNase Inhibitor (RT35), RNase H (RT34), T4 DNA Ligase (EN11), TRANSCRIPTME Reverse Transcriptase (RT32), Tth DNA Ligase (EN13) Year 2020 Source In Situ Hybridization Protocols. Methods in Molecular Biology, vol 2148 -
Hybridization-based in situ sequencing (HybISS) for spatially resolved transcriptomics in human and mouse brain tissue
Authors Daniel Gyllborg, Christoffer Mattsson Langseth, Xiaoyan Qian, Eunkyoung Choi, Sergio Marco Salas, Markus M Hilscher, Ed S Lein, Mats Nilsson Products phi29 Polymerase (EN20), RIBOPROTECT Hu RNase Inhibitor (RT35), TRANSCRIPTME Reverse Transcriptase (RT32), Tth DNA Ligase (EN13) Year 2020 Source Nucleic Acids Research -
TGFβR-SMAD3 Signaling Induces Resistance to PARP Inhibitors in the Bone Marrow Microenvironment
Authors Bac Viet Le, Paulina Podszywalow-Bartnicka, Silvia Maifrede, Katherine Sullivan-Reed, Margaret Nieborowska-Skorska, Konstantin Golovine, Juo-Chin Yao, Reza Nejati, Kathy Q. Cai, Lisa Beatrice Caruso, Julian Swatler, Michal Dabrowski, Zhaorui Lian, Peter Valent, Elisabeth M. Paietta, Ross L. Levine, Hugo F. Fernandez, Martin S. Tallman, Mark R. Litzow, Jian Huang, Grant A. Challen, Daniel Link, Italo Tempera, Mariusz A. Wasik, Katarzyna Piwocka, Tomasz Skorski Products RIBOPROTECT Hu RNase Inhibitor (RT35) Year 2020 Source Cell Reports Volume 33, Issue 1 -
Limited reverse transcriptase activity of phi29 DNA polymerase
Authors Tomasz Krzywkowski, Malte Kühnemund, Di Wu and Mats Nilsson Products Deoxyribonucleotides (dNTPs), RNase Inhibitor Year 2018 Source Nucleic Acids Research, 2018, Vol. 46, No. 7 3625–3632 -
Padlock Probes to Detect Single Nucleotide Polymorphisms
Authors Tomasz Krzywkowski and Mats Nilsson Products TRANSCRIPTME RNA Kit (RT31), RNase H (RT34), RNase Inhibitor Year 2017 Source Springer Science+Business Media LLC 2018 -
In Situ Single-Molecule RNA Genotyping Using Padlock Probes and Rolling Circle Amplification
Authors Tomasz Krzywkowski Thomas Hauling and Mats Nilsson Products RNase H (RT34), TRANSCRIPTME RNA Kit (RT31), RNase Inhibitor Year 2016 Source Springer Science+Business Media New York 2017