Ribonuclease A (RNase A) - DNase-free
Advantages at a glance
- free of DNase activity
- directly usable in DNA purification procedures
- from BSE free sources
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Product information "Ribonuclease A (RNase A) - DNase-free"
Ribonuclease A- DNase-free is a DNase and Protease-free enzyme that can be used directly for RNA digests, e.g. in DNA purification procedures. Elimination of potential DNAse activity is not necessary. Potential DNAse activities have been removed.
Description
Ribonuclease A is an endo-ribonuclease specifically cleaving single stranded RNA at the 3' side of pyrimidine bases (cytosine and uracil). RNAse A ist used to prepare RNA-free DNA, to digest non-hybridised regions of RNA-DNA hybrids and as a molecular weight marker. The pH optimum for RNAse A is between 7.0-7.5. RNAse A is inhibited by Diethylpyrocarbonate (DEPC) >, guanidinium salt (4M GuaSCN), beta-Mercaptoethanol, heavy metals, vanadyl-ribonucleoside complexes, RNAse-inhibitor from human placenta and by competitive DNA. RNAse A cleaves single and douple stranded RNA and RNA in RNA:DNA hybrids at low salt conditions (100mM NaCl). At high salt conditions (>300 mM NaCl), RNAse A cleaves single-stranded RNA only. The enzyme can only be removed by Proteinase K digest and subsequent phenol extraction.
Stock solution: Stock solutions are prepared at concentrations from 1 - 10mg/mL in 10mM Tris/HCl, pH7.5; 15mM NaCl or in 10mM Tris/HCl, pH7.5; 1mM EDTA, pH8.0 (TE buffer).
The recommended working concentration is 10µg/mL (removal of RNA from plasmid preparations; 1hr, RT) or 100ng/mL (preparation of 'blunt ends' of double-stranded cDNA).
Stability:RNase A aggregates during lyophilizing and storage. It has a high affinity to glas surfaces, which has to be taken into consideration. At neutral pH (e. g. in PBS pH 7.4) and high concentrations (>10mg/mL) the enzyme will precipitate. At +2°C to +8°C (lyophilized) it is stable for several years (dry storage), in solution (-20°C) several years or (+2°C to +8°C) several weeks.
Specifications:
Activity: min. 80 units/mg
DNases: not detectable
Proteases: not detectable
MW = approx. 13,700 Da
Product is isolated from BSE free source.
Optimal temperature: 60°C (activity range of 15 - 70°C); Optimal pH: 7.6 (activity range of 6–10); Inhibitors: ribonuclease inhibitor. Activators of RNase A: potassium and sodium salts.
Applikation / Application:
RNase protection assays Remove unspecifically bound RNA Analysis of RNA sequences Hydrolyze RNA contained in protein samples Purification of DNA Pancreatic RNase A specifically cleaves at the 3'-side of pyrimidine (uracil or cytosine) phosphate bondsEinheiten / Units:
1 unit corresponds to the amount of enzyme which hydrolyzes RNA at a rate constant k = 1 at 25°C and pH 5.0 (Kunitz-units), M. Kunitz, J. Biol. Chem. 164, 563 (1946).Quelle / Source:
bovine pancreasKlassifizierungen / Classification
Documents:
CertificateProduct description
Source: NCBI PubMed
Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells
Ieva Bruzauskaite, Jovile Raudoniute, Jaroslav Denkovskij, Edvardas Bagdonas, Sandra Meidute-Abaraviciene, Vaida Simonyte, Daiva Bironaite, Almantas Siaurys, Eiva Bernotiene, Ruta Aldonyte
Cytotechnology. 2017 Feb; 69(1): 1–17.
doi: 10.1007/s10616-016-0021-z
PMCID: PMC5264619
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