RNase A

RNase A

From Eukaryotic cells(Yeast) with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease

Catalog Number: GPE007
CAS No.: 9001-99-4 E.C.: 3.1.27.5
Synonyms: Ribonuclease A, Pancreatic Ribonuclease, Ribonuclease I,  Ribonuclease 3’-pyrimidinooligonucleotidohydrolase

 

Properties
Molecular mass 13.7 kDa
pI 9.6
pH range 6-10
optimal pH 7.6
Storage Temperature 4 °C for lyophilized powder
-20 °C for solution
Form The product is available as lyophilized powder and solution containing 10 mM Tris-HCl, 50% glycerol, pH 8.0
Temperature profile maximum activity at 60°C
Maximum activity observed at 60 °C, although the enzyme exhibit activity in the temperature range of 15-70 °C.
Specific activity ≥ 60 Kunitz units/mg of protein
Unit Definition:

One unit of the enzyme causes an increase of absorbance by 1.0 unit at 260 nm when yeast RNA is hydrolyzed at 37°C and pH 5.0.

Fifty units are approximately equivalent to 1 Kunitz unit.

Extinction coefficient E1% = 7.1 (280 nm)
Purity and Quality
Molecular biology grade

Endonuclease and exonuclease none detected

Free of DNase activity. No need to heat it before use.

Product Description
General Description:

RNase A is an endoribonuclease that specifically cleaves the single strand RNA at the phosphodiester bond between the 5’-ribose of a nucleotide and the 3’ OH phosphate group of an adjacent pyrimidine nucleotide.

The highest activity is demonstrated with single stranded RNA.

Application:

RNase A is used to remove RNA from plasmid and genomic DNA preparation. It also used to remove RNA from protein samples.

RNase protection assays

Analysis of RNA sequences

Mapping single-base mutations in DNA or RNA

Advantages of  Recombinant RNase A

The recombinant enzyme is identical to the native RNase A in amino acid sequence, molecule structure and specifications. However, the recombinant preparation has advantage in lot-to-lot consistency, superior purity and cost-efficiency.

Preparation Notes
Activators Potassium and sodium salts
The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well the RNA strand in RNA-DNA hybrids. However, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA
Inhibitors By alkylation of His 12 and His 119
The RNase A has a high affinity to glass surfaces.

At neutral pH and high concentrations (>10 mg/ml) the enzyme will precipitate.

The enzyme is inhibited by diethyl pyrocarbonate (DEPC), guanidinium salts (4 M GuSCN), β-mercaptoethanol, heavy metals and RNase-inhibitors like RIBOPROTECT (RT33).

In order to remove the enzyme from a sample, perform a separation with spin columns or several phenol/chloroform extractions.

Preparation Instructions
Recommended concentration of RNase A is 1 to 100 µg/mL depending on the application.

For removal of RNA from plasmid preparations use 10 μg/ml working solution and incubate sample for 1 hour at room temperature.

For preparation of „blunt ends” of double-stranded cDNA use 100 ng/ml working solution.

Stability storage at -20 °C
Gproan recommends storage at -20 °C for solution and lyophilized powder to maintain stable for at least 2 years.

RNase A is a very stable enzyme and solutions have been reported to withstand temperatures up to 100 °C. At 100 °C, an RNase A solution is most stable between pH 2.0 and 4.5

Precautions and Disclaimer
This product is for R&D use only, not for drug, household, or other uses.
RNase A is stable to both heat and detergents. In addition, it adsorbs strongly to glass. Scrupulous precautions are necessary to ensure RNase A residue does not cause artifacts in processes requiring intact RNA.
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