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Medical Laboratory Reagents

RNase A
CAS No. 9001-99-4     EC No. 3.1.27.5
From Eukaryotic cells(Yeast) with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease
Catalog/Part No.    CT-RA-037
Synonyms Ribonuclease A, Pancreatic Ribonuclease, Ribonuclease I, Ribonuclease 3-pyrimidinooligonucleotidohydrolase
Properties
Molecular Weight 13.7 KDa
pI 9.6
pH range pH 6-10
optimal pH 7.6
Storage Temperature 4 C for lyophilized powder
-20 C for solution
Form lyophilized powder
Temperature profile maximum activity at 60 C
Temperature range of enzyme activity: 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 37C 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.
Advantages 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.
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
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), -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 g/mL depending on the application.
For removal of RNA from plasmid preparations use 10 g/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
recommended 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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