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RiboLock™ RNase Inhibitor

#EO0381	2500 u (40 u/µl)
#EO0382	4x2500 u (40 u/µl)
#EO0384	24x2500 u (40 u/µl)
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EO0381, #EO0382, #EO0384 MSDS (English) MSDS (English-USA) MSDS (German) Brochure (in pdf, 522 KB)

 

Features

• Performs under a wide range of reaction conditions.
• Protects RNA from degradation at temperature up to 55°C (see Fig.1).

RiboLock™ RNase Inhibitor, brochure in pdf, 522 KB

Description
The RiboLock™ RNase Inhibitor inhibits the activity of RNases A (see Fig.2), B and C by binding them in a noncompetitive mode at a ratio 1:1. It does not inhibit the following RNases: I, T1, T2, H, U1, U2 and CL3.

Source
E.coli cells carrying a cloned gene encoding mammalian ribonuclease inhibitor.

Molecular Weight
49.6 kDa monomer.

Applications

• Inhibition of RNA degradation in the following procedures:
  • in vitro transcription (1);
  • cDNA synthesis (2);
  • in vitro translation (3);
  • isolation of mammalian cell fractions that contain mRNA-protein complex (3).
• Separation and identification of specific ribonuclease activities (4).
• Studies of tumor suppression (5).

Quality Control
The absence of endodeoxyribonucleases, exodeoxyribonucleases, phosphatases and ribonucleases confirmed by appropriate quality tests. Functionally tested in RNA and cDNA synthesis.

Concentration
40 u/µl

Definition of Activity Unit
One unit of the protein inhibits the activity of 5ng RNase A by 50%.
Inhibitor activity is assayed in the following mixture: 100 mM Tris-HCl (pH 7.5), 1.2 mM EDTA, 0.1 mg/ml BSA, 100 ng/ml RNase A, 0.1 mg/ml E.coli [³H]-RNA, 50 mg/ml yeast RNA, 8 mM DTT.

Storage Buffer
The protein is supplied in: 20 mM HEPES-NaOH (pH 7.5), 50 mM NaCl, 8 mM DTT, 0.5 mM ELUGENT Detergent and 50% (v/v) glycerol.

Note

• DTT is required to ensure RiboLock™ RNase Inhibitor stability during long term storage, but it is not necessary for inhibitor activity.
• Working concentration -1 unit per µl of a reaction mixture.

Figure 1. Thermostability of RiboLock™ RNase Inhibitor. Total human RNA (1 µg) was supplemented with 20 u of RiboLock™ RNase Inhibitor and 50 pg of RNase A, DNase and protease-free, and incubated at different temperatures. M - RiboRuler™ RNA Ladder, High Range (#SM1821) C - total human RNA 1 - total human RNA with RNase A 2-6 - total human RNA with RiboLock™ RNase Inhibitor and RNase A

Figure 2. Inhibition of RNase A by RiboLock™ RNase Inhibitor. Total human RNA (1 µg) was supplemented at 37°C with 20 u of RiboLock™ RNase Inhibitor and various amounts of RNase A, DNase and protease-free. M - RiboRuler™ RNA Ladder, High Range (#SM1821) C - total human RNA 1 - total human RNA with RNase A 2-5 - total human RNA with RiboLock™ RNase Inhibitor and RNase A

 

Related Products

RNA Polymerases: SP6 RNA Polymerase T3 RNA Polymerase T7 RNA Polymerase T7 Transcription Kit M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase RevertAid™ H Minus M-MuLV Reverse Transcriptase First Strand cDNA Synthesis Kit RevertAid™ First Strand cDNA Synthesis Kit RevertAid™ H Minus First Strand cDNA Synthesis Kit T4 RNA Ligase DTT DEPC-treated Water

References

1. Nielsen, D.A., Shapiro, D.J., Preparation of capped RNA transcripts using T7 RNA polymerase, Nucleic Acids Res., 14, 5936, 1986.
2. Martynoff, G., et al., Synthesis of a full length DNA complementary to thyroglobulin 33S messenger RNA, Biochem. Biophys. Res. Commun., 93, 645-653, 1980.
3. Scheele, G., Blackburn, P., Role of mammalian RNase inhibitor in cell-free protein synthesis, Proc. Natl. Acad. Sci. USA, 76, 1898-1902, 1979.
4. Eichler, D.C., et al., Effect of human placental ribonuclease inhibitor in cell-free ribosomal RNA synthesis, Biochem. Biophys. Res. Commun., 101, 396-403, 1981.
5. Polakowski, I.J., et al., A ribonuclease inhibitor expresses anti-angiogenic properties and leads to reduced tumor growth in mice, Amer. J. Pathol., 143, 507-517, 1993. 

Updated vasario 25, 2008 10:24