RNase A Powder
BR Grade,For Research Use Only
Cat. No. N9047
Size: 1 g
Specific Activity: ≥3000 U/mg protein (≥60 Kunitz units/mg protein).
Form: Contains 1 g RNase A, lyophilized powder
Description
The RNase A is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. It cleaves the phosphodiester bond between the 5'-ribose of a nucleotide and the phosphate group attached to the 3'-ribose of an adjacent pyrimidine nucleotide. The resulting 2', 3'-cyclic phosphate is hydrolyzed to the corresponding 3'-nucleoside phosphate.
Applications
--Plasmid and genomic DNA preparation
--Removal of RNA from recombinant protein preparations.
--Ribonuclease protection assays
--Mapping single-base mutations in DNA or RNA
Storage
-20ºC recommended. The shelf life of RNase A powder is one year when stored sealed and dry. RNase A powder is shipped at room temperature.
Quality Control
Functionally tested for RNA digestion in a plasmid DNA purification procedure.
Molecular Weight
13.7 kDa monomer.
Definition of Activity Unit
One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm when yeast RNA is hydrolyzed at 37°C and pH 5.0.
Fifty units are approximately equivalent to 1 Kunitz unit.
Inhibition and Inactivation
--Inhibitors: the most potent inhibitor is a ~50 kDa protein from cytosol of mammalian cells, e.g., RiboLock™ RNase Inhibitor.
--Other inhibitors: uridine 2',3'-cyclic vanadate, 5'-diphosphoadenosine 3'-phophate and 5'-diphosphoadenosine 2'-phophate (2), SDS, diethyl pyrocarbonate, 4M guanidinium thyocyanate plus 0.1M 2-mercaptoethanol and heavy metal ions. Inactivated by phenol/chloroform extraction.
--Inactivated by phenol/chloroform extraction.
--Inactivated by heating at 95°C for 10 minutes.
Note
--The working concentration for RNase A is 1-100 µg/ml depending on the application.
--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
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