Ribonuclease Inhibitor, human

Product No. R2520

Product Description

Ribonuclease Inhibitor from human placenta is a protein with a molecular mass of ~50 kDA by SDS-PAGE.1 Inhibition of ribonuclease activity occurs by the formation of a tight, non-covalent 1:1 complex having a dissociation constant (Ki) of 4 x 10–14 M.1-3 Uses include in vitro inhibition of ribonucleases in procedures such as cDNA synthesis from mRNA, in vitro transcription/ translation reactions,3 ribonuclease protection assays,4 and RT-PCR.5 While this Ribonuclease Inhibitor inhibits RNase A, RNase B, and RNase C, it does not inhibit RNase H, S1 Nuclease, SP6, T7 or T3 RNA Polymerase, AMV or M-MLV Reverse Transcriptase, RNase 1, RNase T1, or Taq Polymerase.

The pH range for inhibition is pH 5–9 (highest inhibition at pH 7–8).6 Typical concentration for use is 250–1000 units/ml.7 Denaturing conditions (i.e., urea or temperatures ≥50 °C) should be avoided as they may cause release of active ribonuclease from the complex.

The Ribonuclease Inhibitor may be removed by phenol extraction or inactivated by heating at 65 °C for 10 minutes.

Storage

Storage Temperature –20 °C

Components

This product is isolated from human placenta and supplied in a solution containing 20 mM HEPES-KOH, pH 7.6, with 50 mM KCl, 8 mM DTT, and 50% (v/v) glycerol.

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

Specificity

Specific activity: 30,000–50,000 units/ml.

Unit definition: One unit will cause the inhibition of 50% of the activity of 5 ng of ribonuclease A in a cytidine 2',3'-cyclic monophosphate system.8

Materials

     

 References

  1. Blackburn, P. et al., J. Biol. Chem., 252, 5904 (1977).
  2. Lee, F.S. et al., Biochem., 28, 219 (1988).
  3. Lee, F.S. et al., Biochem., 28, 225 (1989).
  4. Ausubel, F. et al., eds., Short Protocols in Molecular Biology, third edition, John Wiley & Sons, (New York, NY: 1995) p.4-17.
  5. Innis, M.A., and White, T.J., eds., PCR Protocols: A Guide to Methods and Applications, Academic Press, (New York, NY: 1990) p.23 (Catalog Number P8177).
  6. Blackburn, P., and Moore, S., Pancreatic Ribonucleases, in The Enzymes, Vol XV, Part B Academic Press, (New York, NY: 1982).
  7. Farrell, Jr., R.E., RNA Methodologies: A Laboratory Guide for Isolation and Characterization, Academic Press, (New York, NY: 1993) pp. 34-45.
  8. Blackburn, P., J. Biol. Chem., 254, 12484 (1979).
  9. de Martynoff, G. et al., Biochem. Biophys. Res. Commun., 93, 645 (1980).
  10. Scheele, G., and Blackburn, P., Proc. Natl. Acad. Sci. USA, 76, 4898 (1979).

 

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