| RNase A
From Eukaryotic cells（Yeast） with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease |
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| 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. |
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| 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. |
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| 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. |
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| 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. |
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| 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. |
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| 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 |
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| 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. | |
