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Uvod
Characteristics of RNase A:
- Enzyme Type:
- RNase A is an endoribonuclease that specifically degrades single-stranded RNA.
- Specificity:
- It specifically cleaves RNA at cytosine (C) and uracil (U) residues.
- Action Mechanism:
- The enzyme cleaves the phosphodiester bonds between the 5′-ribose of a nucleotide and the phosphate group attached to the 3′-ribose of an adjacent pyrimidine nucleotide.
- This action initially forms a 2′,3′-cyclic phosphate, which is subsequently hydrolyzed to a 3′-nucleoside phosphate.
- Structure:
- RNase A is a single-chain polypeptide that includes 4 disulfide bridges, contributing to its stability.
- Optimal Activity:
- The highest enzymatic activity of RNase A is observed when acting on single-stranded RNA.
Applications of RNase A:
Primary Use:
- RNase A is extensively used to remove RNA from preparations of plasmid DNA, ensuring that the DNA samples are free of RNA contamination.
Activity Across Different Conditions:
Salt Concentration Influence:
- At low salt concentrations (0 to 100 mM NaCl), RNase A can cleave both single-stranded and double-stranded RNA, and the RNA strand in RNA-DNA hybrids.
- At higher salt concentrations (0.3 M NaCl or more), its activity becomes specific to single-stranded RNA.
pojedinosti
Tehnički podaci
| Značajke | Tehnički podaci |
| Appearance | White or light yellow lyophilized powder |
| Molecular weight | 13.7 KDa |
| Purity | ≥60% RNase A (SDS-PAGE) |
| Specific activity | >50 Kunitz units/mg protein |
| Preservation conditions | -20-8 °C. RNase is stable under heating and detergent conditions and can withstand high temperature treatment of 100 °C. |
| DNase residue detection | Not detected. In 10μl (including 200 ng plasmid) solution, RNase A was added to the final concentration of 50-100 μg/ml and placed at room temperature for 30 minuta. After electrophoresis, the main band was obvious without degradation. |
| Application | Remove RNA from preparations of plasmid DNA |
Prednosti
- High activity, >50 U/mg
- High cost performance
- Visoka čistoća, low impurity protein pollution
- No residual Dnase or trace Dnase
Ordering information
| Sadržaj | C12120 | C12121 |
| RNaza A, Lyophilizate, >50 U/mg of protein | 1 g | 10 g |
Skladištenje i stabilnost
Magen recommends storage at -20-8 °C. When stored at 2-8 °C, the product retains activity for at least 2 years.
Recenzije
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