RiboShield™ RNase Inhibitor is a recombinant protein that blocks the activity of a wide range of ribonucleases to reliably protect your RNA from RNase digestion.
The inhibitor is designed for use in RNA-sensitive applications such as RT-qPCR, cDNA synthesis and RNA sequencing, where the presence of even small amounts of RNase can be highly detrimental to RNA quality and experimental outcome.
- Superior protection leading to better performance of RNA-sensitive applications
- Particularly suited to incorporation into saliva-based tests for SARS-CoV-2 detection
- Inhibits eukaryotic RNases, including RNase A, B and C
- Compatible with reverse transcriptases, RNA polymerases and Taq DNA polymerase
- Stable up to 65°C for at least 30 minutes
- Ribonuclease and phosphatase free
- Ideal for long term storage of samples
Ribonucleases (RNases) are found in all cell types and organisms, and their ubiquitous nature can be highly problematic in the lab. Contamination with even the smallest amount of RNase in an RNA sample is enough to completely destroy the RNA. RiboShield™ RNase Inhibitor is a recombinant human placental protein that blocks the activity of a wide range of ribonucleases, including RNase A, B and C, to reliably protect your RNA from RNase digestion.
Superior RNA protection
RiboShield™ RNase Inhibitor is designed for RNA-sensitive applications including RNA isolation, RT-qPCR and RNA sequencing, to shield your RNA from degradation and provide higher yields and better performance as a result.
The inhibitor binds noncovalently to RNases at a 1:1 ratio and with very rapid kinetics of association, guaranteeing immediate protection of your RNA. RiboShield™ RNase Inhibitor has been tested in RT-qPCR and offers the greatest RNA protection when compared to leading competitor products (see figure 1).
Stability at higher temperatures
RiboShield™ RNase Inhibitor is able to perform over a wide range of reaction conditions and can sustain inhibition of RNase A at temperatures up to 65°C for at least 30 minutes (see figure 2). In addition, RiboShield™ does not contain cysteine residues that have been implicated in the oxidation sensitivity of the human placental version of the protein1. This results in an RNase inhibitor molecule that is not only thermostable, but also more resistant to oxidative stress.
The inhibitor can be used to block the activity of a wide range of ribonucleases, including eukaryotic RNases of the neutral type (RNases A, B and C). It does not inhibit RNases T1, T2, U1, U2, CL3, RNase I and H. The inhibitor does not hinder other enzymes such as reverse transcriptases, RNA polymerases or Taq DNA polymerase making it compatible with many enzymatic reactions involving RNA.
RiboShield™ is purified from a strain of Pichia pastoris expressing a modified human placental gene. The inhibitor is free from ribonucleases and phophatases and is inactivated by heating at 75°C for 15 minutes.
1 Kim BM, Schultz LW, Raines RT. Variants of ribonuclease inhibitor that resist oxidation. Protein Science. 1999; 8(2):430-434.
- cDNA synthesis
- 1-step RT-PCR
- 1-step RT-qPCR
- RNA purification
- RNA sequencing
- In vitro transcription and translation
- Saliva-based diagnostic testing for SARS-CoV-2
The kit should be stored between -30°C and -15°C. Avoid prolonged exposure to light. If stored correctly the kit will retain full activity for 12 months. The kit can be stored at 4°C for 1 month. The kit can go through 30 freeze/thaw cycles with no loss of activity.
Superior Protection against RNase A in RT-qPCR:
10 different dilutions of RiboShield RNase Inhibitor and each of three competitor products were incubated with 5ng RNase A and 2ug RNA in 5x UltraScript buffer at 37°C for 30 min. After this step, RNA samples were reverse transcribed using UltraScript Reverse Transcriptase with oligo(dT) primers.
Amplification of g-Actin was performed using qPCRBIO SyGreen Blue Mix No-ROX. The RNase inhibitors used were PCR Biosystems RiboShield, Promega’s RNasin, Bioline’s RiboSafe and ThermoFisher’s RNaseOUT. Internal standards on each plate were used to convert Ct values into ug RNA n=4. Data are plotted as amount of reverse transcribable RNA per concentration of RNase inhibitor used, in a linear (a) and logarithmic (b) scale. IC50 values were calculated for each of the RNase inhibitors used.
RiboShield has the lowest IC50 value and offers the greatest RNA protection amongst the inhibitors tested.
Stable at High Temperatures:
RiboShield RNase inhibitor was incubated at the indicated temperatures for 30 minutes. 40U of the inhibitor were then added to 1ug RNA and 5pg RNase A in 5x UltraScript buffer and incubated at 37°C for 30 min. Samples were then loaded on a 1% agarose gel.
RiboShield RNase Inhibitor can inhibit RNase A at temperatures up to 65°C for at least 30 minutes.
Superior Protection Against RNase A:
RiboShield RNase Inhibitor and three competitor products (40U) were incubated with the indicated amounts of RNase A and 1ug RNA in 5x UltraScript buffer at 37°C for 30 minutes. Samples were then loaded on a 1% agarose gel L.Ambion RNA Millenium Markers. The RNase inhibitors used were PCR Biosystem’s RiboShield, Promega’s RNasin, Bioline’s RiboSafe and ThermoFisher’s RNaseOUT.
RiboShield offers the greatest RNA protection amongst the tested inhibitors.
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|RiboShield™ RNase Inhibitor (40U/µL)||
1 x 62.5µL | 2500 Units
4 x 62.5µL | 10000 Units