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Analysis Note

Protein determined by E.

Application

Ribonuclease A from bovine pancreas has been used:for plasmid DNA purificationto digest and avoid staining of double-stranded RNA during staining of nucleiin immunoperoxidase and immunofluorescence detectionto treat cells prior to flow cytometryas a component of post-hybridization buffer for washing FFPE tissue sections post in-situ hybridizationto treat cells during the preparation of outer membrane protein (OMP) extractsRNase protection assaysRemoval of unspecifically bound RNAAnalysis of RNA sequencesHydrolysis of RNA contained in protein samples

RNase A is used to remove RNA from DNA plasmid and genomic DNA preparations and protein samples. RNase A is also used in RNA sequence analysis and protection assays. RNase A has been used as a tool for computer-aided drug design. RNase A supports the analysis of RNA sequences. RNase A hydrolyze RNA contained in protein samples. Purification of DNA is supported by RNase A.

Features and Benefits

Our highly stable Ribonuclease A, RNase A, is suitable for removal of RNA, RNA sequencing, and DNA purification.

General description

RNase A, Ribonuclease A, is an endoribonuclease that cleaves the phosphodiester bonds of single strand RNA after pyrimidine nucleotides. It attacks at the 3′ phosphate end (For example pG-pG-pC-pA-pG will be cleaved to give pG-pG-pCp and A-pG). The highest activity is exhibited with single stranded RNA. RNase A is a single chain polypeptide containing 4 disulfide bridges. In contrast to RNase B, it is not a glycoprotein. Ribonucleases do not hydrolyze DNA, because the DNA lacks 2′-OH groups essential for the formation of cyclic intermediates. RNase A can also hydrolyze RNA from protein samples. RNase A can be inhibited by alkylation of His12 and His119 and activated by potassium and sodium salts. RNAse is inhibited in the presence of heavy metal ions. RNase is also inhibited competitively by DNA.

RNase A is a single chain polypeptide containing 124 amino acids, linked by four disulfide bridges. In contrast to RNase B, RNase A is not a glycoprotein. RNase A can be inhibited by alkylation of His¹² or His¹¹⁹, which are present in the active site of the enzyme. Activators of RNase A include potassium and sodium salts.

RNase A is an endoribonuclease that attacks at the 3’;OHphosphate of a pyrimidine nucleotide. The sequence of pG-pG-pC-pA-pG will be cleaved to give pG-pG-pCp and A-pG. The highest activity is exhibited with single stranded RNA.

Other Notes

A major application for RNase A is the removal of RNA from preparations of plasmid DNA. For this application, DNase free RNase A is used at a final concentration of 10 µg/mL.Boiling stock solutions of this RNase A product to inactivate residual DNase is not necessary and may cause precipitation of RNase and possible loss of enzymatic activity. If an RNase A solution is heated at a neutral pH, precipitation will occur. When heated at a lower pH, some precipitation may occur because of protein impurities that are present.

Activators of RNase A include potassium and sodium salts. RNase A can be inhibited by alkylation of His12 or His119.