RNase A is an endoribonuclease that attacks at the 3'-phosphate of a pyrimidine nucleotide. The sequence of pG-pG-pC-pA-pG will be cleaved to give pG-pG-pCp and A-pG. The highest activity is exhibited with ssRNA.
RNase A is free of detectable DNase and protease activity, a heat treatment of the enzyme is not necessary before use.
Shipping: Shipped at ambient temperature
Storage Conditions: Store at 2-8 °C. Stock solutions in TE buffer should be aliquoted and stored at -20 °C
Shelf Life: 12 months
Molecular Weight: 13.7 kDa (monomer)
CAS#: 9001-99-4
EC number: 232-646-6
Purity: ≥ 90 % (ion exchange chromatography), salt free, chromatographically homogeneous lyophilisate
Form: dry powder
Activity: ≥ 80 Kunitz units/mg
Unit definition: 1 Kunitz unit is that amount of activity which is capable of causing within 1 minute a decrease in absorbance at
300 nm equivalent to the maximum possible change in a 0.05 % solution of yeast RNA at 25 °C, pH 5.0.
Applications:
- Plasmid and genomic DNA preparation
- Removal of RNA from recombinant protein preparations
- Ribonuclease protection assays
- Mapping single-base mutations in DNA or RNA
Reaction conditions:
Working concentration: 1 - 100 μg/ml (depending on application)
The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase cleaves ss and dsRNA as well as the RNA strand in RNA-DNA hybrids. At NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves ssRNA.
Stability:
RNase A is an extremely stable enzyme, remarkable resistant to heating. It renatures easily after treatment with most denaturing agents.
Inactivation:
Ribonuclease inhibitor, Vanadyl-ribonucleoside complexes, arabinonucleosides, Zn2+, Cu2+, penicillin, Vitamin B12, SDS, DEPC, 4 M guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol. Most polyanions show some inhibitory effect. Inactivated by phenol/chloroform extraction.
Isoelectric point (pI):9.6
Optimal pH: 7.0 (activity range 6 - 10)
