- Product Name:
- Type:Molecular Diagnostics
- Description:RNase Inhibitor specifically inhibits RNases A, B and C. It inhibits RNases by binding noncovalently in a 1:1ratio with high affinity. It is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase fromAspergillus. In addition, no inhibition of polymerase activity is observed when RNase Inhibitor is used with Taq DNA Polymerase, AMV or M-MuLV Reverse Transcriptases, or Phage RNA Polymerases (SP6, T7, or T3).
The 50 kDa protein inhibits RNases by binding noncovalently in a 1:1 ratio with an association constant greater than 1014.
Protocol for Avoiding RNase Contamination using RNase InhibitorAdd inhibitor to achieve final concentration of 1U/ul in your reaction. During assembly of a reaction, RNaseInhibitor should be added before other components that are a possible source of RNase contamination (i.e.
enzymes, plasmid from a mini prep.)
*RNase Inhibitor is a 50 kDa protein that will be inactivated in denaturing conditions.
**RNase Inhibitor should be used below 50°C.
20 mM HEPES-KOH, 50 mM KCl, 8 mM DTT, 50% Glycerol, pH7.6
RNase Inhibitor is tested for contaminating activities, with no trace of endonuclease activity, nicking activity or exonuclease
Since ribonucleases typically retain activity under denaturing conditions, care must be taken to avoid denaturing RNase Inhibitor molecules which have complexed with a ribonuclease. To prevent the release of active ribonuclease, temperatures greater than 50°C and high concentrations of urea or other denaturing agents should be avoided.The recommended concentration of RNase Inhibitor in a reaction is 1 U/μl. During assembly of a reaction,RNase Inhibitor should be added before other components that are a possible source of RNase contamination (i.e. enzymes, plasmid from a mini prep.)
- Storage:Long term storage at -20 °C, short term storage at 2-8 °C
- Source:Recombinant in E.coli
- Active:40 U/μL
- Activity is defined:One unit is defined as the amount of RNase Inhibitor, Mammalian required to inhibit the activity of 5ng of RNase A by 50%. Activity is measured by the inhibition of hydrolysis of cytidine 2', 3'-cyclic monophosphate by RNase A.
- Application:cDNA synthesis, In vitro translation/transcription, RT-PCR
- Size:2000 U or user-defined