Science Catalog 2012 Life Cell Signaling Ribonuclease Inhibitors RNasin® Plus RNase Inhibitor Product RNasin® Plus RNase Inhibitor For Laboratory Use. Description: 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 is 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. The 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. We have taken solutions up to 70°C for 15 minutes and did not see RNase reactivation. Features: • Improved Resistance to Oxidation: Due to natural amino acid diversity, RNasin® Plus lacks the capability to form the active site-blocking disulfide bond that can form in the human protein under oxidative conditions. • Improved Purification: RNasin® 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 >90% pure protein with no E. coli RNase carryover. • Proven Compatibility with RT-PCR Systems: RNasin® Plus has proven compatible with the Access and AccessQuick™ RT-PCR Systems, ImPromII™ Reverse Transcription System and the Reverse Transcription System. Also proven compatible with TaqMan®-based RT-PCR Systems. • Protection During RNA Template Denaturation: Heating mixtures of RNasin® Plus 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® Plus at this step. • Protection During Higher Temperature RT Reactions: Add RNasin® Plus during RT reaction assembly and take the reaction to temperatures above 50°C with enzymes like the ImProm-II™ and AMV Reverse Transcriptases. RNases that may be present will not be reactivated at the higher temperature. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –20°C. Protocol RNasin® Plus RNase Inhibitor Product Information Part# 9PIN261 Size Conc. Cat.# Price (Fr) 2,500 u 40 u/µl N2611 186.00 10,000 u 40 u/µl N2615 465.00 Product RNasin® Ribonuclease Inhibitors Size RNasin® Ribonuclease Inhibitor Recombinant RNasin® Ribonuclease Inhibitor Conc. Cat.# Price (Fr) 2,500 u 20–40 u/µl N2111 218.00 10,000 u 20–40 u/µl N2115 545.00 2,500 u 20–40 u/µl N2511 169.00 10,000 u 20–40 u/µl N2515 423.00 N2511, N2515 For Laboratory Use. N2111, N2115 For Research Use Only. Not for Use in Diagnostic Procedures. Description: Natural and Recombinant RNasin® Ribonuclease Inhibitors have broad-spectrum RNase inhibitory properties, including the inhibition of eukaryotic RNases of the neutral type. The 50kDa protein exerts its inhibitory effect by noncovalently binding to RNases in a 1:1 ratio. The Ki value for the binding of RNasin® Ribonuclease Inhibitor to RNase (e.g., RNase A) is approximately 10–14M. Promega offers two different preparations: Recombinant RNasin® Ribonuclease Inhibitor and Natural RNasin® Ribonuclease Inhibitor. Both products are purified using a combination of ion exchange and affinity chromatography. Features: • Inhibits Common Eukaryotic RNases: RNasin® Ribonuclease Inhibitor has broad-spectrum RNase inhibitory properties, including RNase A, RNase B, RNase C and human placental RNase. Does not inhibit RNase T1, S1 nuclease, RNase from Aspergillus, RNase H, RNase ONE™ Ribonuclease. • Compatibility: RNasin® Ribonuclease Inhibitor does not inhibit SP6, T7 or T3 RNA Polymerase; ImProm-II™, AMV or M-MLV Reverse Transcriptase; or Taq DNA polymerase. • Broad pH Range: Active over a broad pH range (pH 5–8). • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –20°C. Protocol RNasin® Ribonuclease Inhibitor Product Information RNasin® Ribonuclease Inhibitor, Recombinant Protocol Part# 9PIN211 9PIN251 See figure on next page for more information. 96 For complete and up-to-date product information visit: www.promega.com/catalog Pagina 99
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