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 polymerase or reverse transcriptase activity
- Free of DNase and RNase activity
- Stable up to 58°C and 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)
- RNA isolation and purification
- cDNA synthesis, RT-PCR, RT-qPCR
- in vitro transcription and translation
- RNase-free monoclonal antibody preparation
20 mM HEPES-KOH (pH 7.6); 50 mM KCl; 8 mM DTT; 50% (v/v) glycerol
- 0.5 – 1 mM DTT presence is essential for optimal activity of the RIBOPROTECT HuRNase 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 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 Hufor 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.
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%.
Store in a freezer without a defrost cycle. The product is stable for 12 months providing it is stored and used correctly.
Shipping on blue ice.
12 months from the date of purchase
Manufacturer: BLIRT S.A.