| RNase A, Animal-Free From Yeast with cloned gene encoding genetically engineered Bovine pancreatic ribonuclease | |
| Catalog Number: GPE007001 | |
| CAS No.: 9001-99-4 | E.C.: 3.1.27.5 |
| Synonyms: Ribonuclease A, Pancreatic Ribonuclease, Ribonuclease I, Ribonuclease 3’-pyrimidinooligonucleotidohydrolase | |
| Properties | |
| Molecular Weight | 14.3 kDa |
| pH range | 6-10 |
| Optimal pH 7.6 | |
| Storage Temperature | -20 °C for lyophilized powder |
| Form | The product is available as lyophilized powder and solution in 10 mM Tris-HCl, 20mM MgCl2, 50% glycerol, pH 8.0 |
| Temperature profile | Maximum activity at 60°C |
| Exhibit activity in the temperature range of 15-70 °C. | |
| Specific Activity | ≥ 40 Kunitz units/mg |
| 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: ≥90% (SDS-PAGE) DNA and RNA none detected Free of Endotoxin Free of DNase activity. No need to heat it before use. Bioburden: Microorganisms count <5 cfu/g | |
| 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 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 advantages: Contaminant-Free: Recombinant RNase A is produced in a controlled environment, significantly reducing the risk of contamination with other nucleases or proteases. Lot-to-Lot Consistency: Recombinant production ensures consistent enzyme activity and quality across different batches, providing reliable performance in experimental and industrial applications. No Endogenous Nuclease Contamination: Since recombinant RNase A is expressed in a controlled system, it is free from DNA contamination, making it ideal for applications such as RNA preparation and analysis, where the presence of DNA could compromise results. Efficient RNA Digestion: Recombinant RNase A retains high specific activity, efficiently degrading RNA without affecting DNA, which is particularly important in DNA purification protocols. Cost-Effectiveness: Recombinant techniques allow for large-scale production, which can reduce costs and make the enzyme more accessible for various applications. Animal-Free Production: Recombinant RNase A is produced without the use of animal-derived materials, making it suitable for laboratories and industries that prioritize ethical sourcing of materials. Application: RNase A is a versatile enzyme used in various applications across molecular biology, biochemistry, and biotechnology. RNA Removal in DNA Purification & Plasmid Isolation: RNase A selectively degrades RNA, leaving the DNA intact and ensuring the purity of the DNA sample for downstream applications. RNA Sequencing and Analysis: Ribonuclease Protection Assays (RPA): The enzyme digests any unprotected RNA, allowing for precise quantification of target RNA. RNA Mapping: It can be employed to map the structure of RNA by selectively degrading single-stranded regions, helping in the study of RNA secondary structures. Protein and Enzyme Preparation: In the preparation of certain proteins or enzymes from biological samples, RNase A is used to remove RNA contaminants that could interfere with protein or enzyme activity assays. RNA Cleanup in Cell and Tissue Culture: RNase A is used in cell and tissue culture to degrade extracellular RNA that could interfere with experimental outcomes or contaminate samples during cell lysis. RNA Degradation in Diagnostic Tests: In some diagnostic assays, particularly those involving nucleic acid detection, RNase A is used to remove RNA to prevent false-positive results caused by RNA contamination. Selective RNA Degradation in In Situ Hybridization: In situ hybridization techniques use RNase A to selectively degrade RNA in tissue sections or cells, allowing for the specific detection of DNA sequences without interference from RNA. RNA Gel Electrophoresis: RNase A is used to degrade unwanted RNA in samples before running gel electrophoresis, ensuring clear separation and analysis of the desired nucleic acids. RNA Footprinting in Study of RNA Structure and Function: RNase A can be used in RNA footprinting experiments to study the interaction of RNA with proteins or other molecules by selectively cleaving accessible regions of RNA. Environmental and Industrial Applications: RNase A can be used in certain bioremediation processes where the degradation of RNA is necessary, such as in the treatment of RNA-containing waste in industrial settings. | |
| Preparation Notes | |
| Activators | 20mM MgCl2 |
| 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. Gproan RNase A demonstrates peak performance when operating in the presence of 20mM MgCl2. | |
| 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. Dilution buffer: 10 mM Tris-HCl, 20mM MgCl2, pH 8.0 Storage buffer: 10 mM Tris-HCl, 20mM MgCl2, 50% glycerol, pH 8.0 | |
| Stability | storage at -20 °C |
| Gproan recommends storage at -20 °C for solution and lyophilized powder to maintain stable for at least 3 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. | |
