Thermo Scientific™ RiboLock RNase Inhibitor (40 U/μL)

Description

RiboLock RNase Inhibitor inhibits the activity of RNases A , B, and C by binding them in a noncompetitive mode at a 1:1 ratio. It does not inhibit eukaryotic RNases T1, T2, U1, U2, CL3 as well as prokaryotic RNases I and H.

• Does not inhibit eukaryotic RNases T1, T2, U1, U2, CL3 as well as prokaryotic RNases I and H
• Performs under a wide range of reaction conditions
• Protects RNA from degradation at temperatures up to 55°C

• The absence of ribonucleases, endo-, exodeoxyribonucleases and phosphatases and ribonucleases confirmed by appropriate quality tests
• Functionally tested in RNA and cDNA synthesis

• E.coli cells with a cloned gene encoding mammalian ribonuclease inhibitor

• 49.6kDa monomer Definition of Activity Unit:
  • One unit of the RiboLock RNase Inhibitor inhibits the activity of 5ng RNase A by 50%
  • Inhibitor activity is assayed in the following mixture: 100mM Tris-HCl (pH 7.5), 1.2mM EDTA, 0.1mg/mL BSA, 100ng/mL RNase A, 0.1mg/mL [³H]-RNA, 50mg/mL yeast RNA, 8mM DTT
  Storage Buffer:
  • The enzyme is supplied in: 20mM HEPES-NaOH (pH 7.5), 50mM NaCl, 8mM DTT, 0.5mM ELUGENT Detergent and 50% (v/v) glycerol
  Inhibition and Inactivation
  • Inhibitors: common denaturants (SDS, urea and all oxidizing reagents (p-chloromercuribenzoate, dissolved oxygen, ions in their higher oxidation states) strongly inhibit RiboLock RNase Inhibitor and release the RNase bound
  • Inactivated by heating at 75°C for 10 min. Residual activity detectable after 10 min. heating at 70°

  Recommended for:

  • Inhibition of RNA degradation in the following: in vitrotranscription (1), cDNA synthesis (2), in vitrotranslation (3), isolation of mammalian cell fractions that contain mRNA-protein complex
  • RNA purification and storage
  • Separation and identification of specific ribonuclease activities
  • Studies of tumor suppression

  Note:

  • DTT provided in the Storage Buffer ensures stability during long term storage, but is not necessary for inhibitor activity
  • Recommended concentration 1u/μl of a reaction mixture