From Eukaryotic cells（Yeast） with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease
|Catalog Number: GPE007|
|CAS No.: 9001-99-4||E.C.: 220.127.116.11|
|Synonyms: Ribonuclease A, Pancreatic Ribonuclease, Ribonuclease I, Ribonuclease 3’-pyrimidinooligonucleotidohydrolase|
|Molecular mass||13.7 kDa|
|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|
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.
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.
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.
|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.
|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.|