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M0307L |
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10,000 units
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40,000 units/ml |
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M0307S |
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2,000 units
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40,000 units/ml |
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- Isolated from a recombinant source
- Inhibits common eukaryotic RNases
- Compatible with Taq Polymerase, AMV or M-MuLV Reverse Transcriptases
- Active over a broad pH range (pH 5-8)
Description:
RNase Inhibitor is a recombinant human placental protein which specifically inhibits ribonucleases (RNases) A, B and C (1). It is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. 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 (2).
Source:
An E. coli strain that carries the Ribonuclease Inhibitor gene from human placenta.
Reaction & Storage Conditions
Unit Definition:
One unit is defined as the amount of RNase Inhibitor required to inhibit the activity of 5 ng of RNase A by 50%. Activity is measured by the inhibition of hydrolysis of cytidine 2, 3'-cyclic monophosphate by RNase A.
Concentration:
40,000 units/ml
Storage Conditions:
20 mM HEPES-KOH
50 mM KCl
8 mM DTT
50% Glycerol
pH 7.6 @ 25°C
Storage Temperature:
-20°C
Notes
Usage notes:- 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.
- As an alternative, the new Murine Rnase Inhibitor offers improved resistance to oxidation and activity at low DTT concentrations, and 50% more units for same price.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Endonuclease Activity:
Incubation of 200 units of RNase Inhibitor with supercoiled plasmid produced no nicked molecules after a two hour incubation at 37°C as determined by gel electrophoresis.
Ribonuclease Assay:
Incubation of 200 units of RNase Inhibitor with 1 µg of RNA at 37°C for 1 hour resulted in no detectable degradation of RNA as determined by gel electrophoresis.
DNase Assay:
Incubation of 200 units of RNase Inhibitor for 1 hour at 37°C with 50 ng of radiolabeled DNA released < 3% of the radioactivity.
References
- Blackburn, P. and Moore, S. (1982) Pancreatic Ribonucleases. The Enzymes, XV, Part B, Academic Press, NY.
- Blackburn, P., Wilson, G. and Moore, S. (1977) J. Biol. Chem., 252,5904.
Companion Products
HiScribe RNAi Transcription Kit
Murine RNase Inhibitor
ProtoScript® First Strand cDNA Synthesis Kit
ShortCut® RNAi Kit
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