Applications for RNase inhibitor:
- Inhibits common eukaryotic RNases
- Active over a broad pH range (pH 5-8)
- high levels of inhibition over a wide range of conditions
RNase Inhibitor is a recombinant human placental protein which inhibits ribonucleases (RNases) A, B and C. It does nit inhibit RNase 1, RNase T1, S1
Nuclease, RNase H or RNase from Aspergillus. There is no inhibition of polymerase activity when the protein is used with Taq DNA Polymerase, AMV or M-MuLV Reverse
Transcriptases, or Phage RNA Polymerases (SP6, T7, or T3).
Concentration: 40 u/µl
20 mM HEPES-KOH (pH7.6), 50 mM KCl, 8 mM DTT and 50% glycerol
Unit definition RNase inhibitor:
One unit is the amount of enzyme required to inhibit by 50% the activity of 5 ng of RNase A at 250C (This inhibitor activity is determined by its ability to inhibit hydrolysis of cyclic 2’,
3’-CMP by RNase A).
Usage or RNase inhibitor:
We recommend to use 1 unit per reaction unit
- Ribonoclease Inhibitor requires at least 1 mM DTT to be active
- Enzyme inhibits in a wide pH range, but most strongly at pH 7 - 8
- avoid temperatures above 50 °C and high concentrations of urea or other denaturing agents
Transportation: RNase inhibitor is shipped on blue ice
Storage: at -20°C for 24 months
Endonuclease: Contains no detectable endonuclease activity. Incubation of 200 units of enzyme with supercoiled plasmid produced no nicked
molecules after a two hour incubation at 37°C as determined by ethidium-stained agarose gel electrophoresis.
Ribonuclease: No ribonuclease activity is observed after 1 µg of RNA is incubated with 200 units of enzyme for 60 minutes at 37°C. The RNA is electrophoresed on an agarose gel and
stained with ethidium bromide. No latent ribonuclease activity is observed after 1 µg of RNA is incubated with 200 units of pre-heated enzyme for 60 minutes at 37°C. The RNA is electrophoresed on
an agarose gel and stained with ethidium bromide.
DNase: 50 ng of radiolabelled DNA is incubated with 200 units of enzyme for 60 minutes at 37°C, and the release of radiolabelled nucleotides is
monitored by scintillation counting of TCA-soluble material. Minimum passing specification is <3% release of input radioactivity into TCA-soluble material.