Product details

Concentration: 10 mg/ml    Size: 1 ml               
 
Specific Activity: >2500 u/mg protein (>50 Kunitz units/mg protein).
Store at -20℃.
For research use only
 
Description:
The RNase A, DNase and protease-free 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
Source:
Bovine pancreas. 

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.
 
Storage Buffer:
The enzyme is supplied in: 50 mM Tris-HCl (pH 7.4) and 50% (v/v) glycerol.
 
Quality Control:
The absence of endodeoxyribonucleases, exodeoxyribonucleases and proteases confirmed by appropriate quality tests. Functionally tested for RNA digestion in a plasmid DNA purification procedure.

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.

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.