The RIBOPROTECT RNase Inhibitoris a recombinant protein isolated and purified from Escherichia coli. It inhibits ribonuclease (RNase) activity enzymes such RNase A, RNase B, RNase C by non-covalent binding in a 1:1 ratio. RIBOPROTECT is intended for use in applications where the presence of RNases may presents 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 shows no activity towards RNase 1, RNase T1, RNase T2, S1 nuclease and RNase H.
Features and advantages
- Completely inhibits RNase A, B and C activity
- No polymerase or reverse transcriptase activity
- Dnase-, RNase- and DNA-free
- Protects RNA at temperatures up to 55 oC
- Stable at a wide temperature range, pH (pH 5-8) and DTT concentration ranges
- Compatible with the TRANSCRIPTME Reverse Transcriptase
- RNA isolation and purification
- cDNA synthesis, RT-PCR, RT-qPCR
- in vitro transcription and translation
- RNA amplification
20 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.5 mM EDTA, 8 mM DTT, 50% (v/v) glycerol
- 0.5-1 mM DTT presence is essential for optimal activity of the RIBOPROTECT RNase Inhibitor
- The storage buffer contains 8 mM DTT, however, if the ratio of the inhibitor to the final sample volume is less than 1:8, then the addition of DTT to a final concentration of 0.5-1 mM DTT is recommended.
- The optimal recommended final concentration of the RIBOPROTECT 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 2 - 4 U/ µl.
- For a standard reverse transcription reaction, use 1 µl (40U) of the RIBOPROTECT inhibitor for a final sample volume of 20 µl.
- For optimal RIBOPROTECT activity,a final DTT concentration of 0.5-1 mM is essential.
- Adding the RIBOPROTECT inhibitor to the reaction before the reagents, which may cause contamination, is recommended.
- Using RIBOPROTECT does not exclude RNase H treatment after amplification of the first strand cDNA synthesis.
The absence of DNase and RNase activity has been confirmed using the relevant procedures. No DNA contamination has been detected. In addition, the functional quality is tested by RT and subsequent PCR and qPCR assays.
One unit is defined as the amount of enzyme required to inhibit the activity of 5 ng RNazy A by 50%. Activity is measured by the inhibition of the hydrolysis of cytidine 2', 3'-cyclic monophosphate by RNase A.
Manufacturer: BLIRT S.A.