| Species : |
Bovine |
| Source : |
Yeast |
| Tag : |
Non |
| 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. |
| Form : |
The product is available as lyophilized powder and solution in 10 mM Tris-HCl, 20mM MgCl2, 50% glycerol, pH 8.0 |
| Bio-activity : |
≥ 40 Kunitz units/mg |
| Molecular Mass : |
14.3 kDa |
| Purity : |
≥ 90% (SDS-PAGE) |
| Applications : |
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. |
| Storage : |
At -20 centigrade for lyophilized powder.
Recommends storage at -20 centigrade 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 centigrade. At 100 centigrade, an RNase A solution is most stable between pH 2.0 and 4.5. |
| Concentration : |
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. |
| CAS No. : |
9001-99-4 |
| 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 centigrade and pH 5.0.
Fifty units are approximately equivalent to 1 Kunitz unit. |
| EC No. : |
3.1.27.5 |
| pH range : |
6-10, Optimal pH 7.6 |
| Bioburden : |
Microorganisms count: < 5 cfu/g |
| Advantages : |
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. |
| Dilution Buffer : |
10 mM Tris-HCl, 20mM MgCl2, pH 8.0 |
| Storage Buffer : |
10 mM Tris-HCl, 20mM MgCl2, 50% glycerol, pH 8.0 |
| Temperature profile : |
Maximum activity at 60 centigrade
Exhibit activity in the temperature range of 15-70 centigrade. |
| 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.
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. |
| Extinction Coefficient : |
E1% = 7.1 (280 nm) |
