RNases are ubiquitous and can cause RNA degradation and compromise RNA integrity. Native and Recombinant RNasin Inhibitors are 50kDa proteins that inhibit RNase A family and human placental RNases by noncovalently binding to RNases in a 1:1 ratio. For downstream applications such as GoScript Reverse Transcriptase, AMV/M-MLV reverse transcriptases, SP6, T7/T3 RNA polymerase, and Taq DNA polymerases, Recombinant RNasin Inhibitor does not inhibit RNase T1, S1 nuclease, RNase from Aspergillus, RNase H, RNase ONE Ribonuclease and enzymes. RNasin Plus RNase Inhibitor is a recombinant mammalian RNase inhibitor that is expressed as a soluble protein in E. coli, allowing easy purification through a combination of ion exchange and hydrophobic interaction chromatography. The protein is capable of inhibiting eukaryotic RNases (e.g., RNase A and RNase B) similarly to human placental RNase inhibitor. RNasin Plus RNase Inhibitor is tested in RT-PCR and compatible with enzymes such as AMV, M-MLV and ImProm-II Reverse Transcriptases or Taq and Tfl DNA Polymerases. RNasin Plus RNase Inhibitor also is tested and compatible with quantitative, real-time RT-PCR in a TaqMan assay. RNasin Plus RNase Inhibitor offers increased resistance to oxidation over the human version of the protein. Two cysteines in the human protein have been identified as especially sensitive to oxidation and react by forming a disulfide bond that can block the active site of the inhibitor. RNasin Plus, through natural amino acid diversity, lacks the ability to form this site-blocking disulfide. In addition, the new protein has characteristics never before realized, including continued inhibition of RNases above 50°C. Heating solutions of RNasin Plus and RNase followed by cooling does not result in the reappearance of RNase activity—even when the solution is heated above the denaturation temperature of the RNasin Plus protein alone. This allows RNasin Plus to protect RNA species prior to, during and after heating, even at temperatures normally used during first-strand DNA synthesis in RT-PCR. Solutions heated up to 70°C for 15 minutes did not result in RNase reactivation.|
Achieve Broad-Spectrum RNase Inhibition: Inhibits common eukaryotic RNases.
Use with Many Enzymes: Does not inhibit SP6, T7 or T3 RNA Polymerase; GoScript(TM); Reverse Transcriptase, AMV or M-MLV Reverse Transcriptase; or Taq DNA polymerase.
Use in Many Downstream Assays: Functional across wide pH range (pH 5-8).
Choose Native or Recombinant Form: Recombinant form is made in bacteria, minimizing the chances of human nucleic acid contamination.
RNasin(R), Plus RNase Inhibitor also can:
Improve Resistance to Oxidation: Due to natural amino acid diversity, RNasin(R), Plus lacks the capability to form the active site-blocking disulfide bond that can form in the human protein under oxidative conditions.
Improve Purification: RNasin(R), Plus is expressed by E. coli as a soluble protein, allowing easy purification by a combination of ion exchange and hydrophobic interaction chromatography. No direct affinity chromatography required. The new process yields a more than90% pure protein with no E. coli RNase carryover.
Use with RT-PCR Systems: RNasin(R), Plus has proven compatible with the Access and AccessQuick(TM); RT-PCR Systems, M-MLV Reverse Transcriptase, ImProm-II(TM); Reverse Transcription System and the GoScript(TM); Reverse Transcription System. Also proven compatible with TaqMan(R),-based RT-PCR Systems.
Protect During RNA Template Denaturation: Heating mixtures of RNasin(R), Plus RNase Inhibitor and RNase does not lead to reactivation of the RNase at temperatures even as high as 70 C for 15 minutes. Many RT-PCR protocols call for RNA template denaturation (e.g., 65-70 C for 5-10 minutes) in the presence of the RT primers prior to full RT reaction assembly for maximum sensitivity. You can now include RNasin(R), Plus at this step.
Protect During Higher Temperature RT Reactions: Add RNasin(R), Plus RNase Inhibitor during RT reaction assembly and take the reaction to temperatures above 50 C with enzymes like the ImProm-II(TM); and AMV Reverse Transcriptases. RNases that may be present will not be reactivated at the higher temperature.