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 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. |
| 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), ß-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
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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