Science Catalog 2012 Life Cell Signaling Product RNasin® Ribonuclease Inhibitors Size Conc. RNasin® Ribonuclease Inhibitor Recombinant RNasin® Ribonuclease Inhibitor A 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 B 0.05 0.1 0.15 0.2 0.25 0 pre-incubation in assay 10 20 30 40 50 60 70 80 90 0 Amount and Type of Inhibitor Added Comparison of RNasin® Ribonuclease Inhibitor and Superase•In™ inhibition of RNase A activity. Panel A. Total yeast RNA assay. Total yeast RNA was incubated in the presence of 5ng RNase A for 5 minutes at 37°C in 0.5ml of reaction mix containing 50mM MOPS and 5mM MgCl2 (pH 6.5). The indicated amounts of inhibitor (RNasin® or Superase•In™) were mixed with the RNA prior to RNase addition. After incubation, 0.5ml 10% TCA was added to stop the reaction and to precipitate the large RNA molecules. An OD280 measurement was taken of the TCA-soluble material. Panel B. “Pre-incubation” and “in assay” conditions. The total yeast RNA assay was performed as described in Panel A along with an experimental modification of “pre-incubation.” For the pre-incubation assay, the ribonuclease inhibitors were mixed with RNase and incubated for 15 minutes at 22°C. The pre-incubation mix was then added to the RNA. 130 For complete and up-to-date product information visit: www.promega.com/catalog RNase A Activity (units) Percent Inhibition of RNase Activity 3109MA10_0A 3074MA09_0A 4u RNasin® 4u Superase•In™ 40u Superase•In™ 400u Superase•In™ RNase A RNase A + 8u RNasin® Ribonuclease Inhibitor 8u Superase•In™ RNase A + 40u Superase•In™ RNase A + Pagina 133
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