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R6513

Sigma-Aldrich

Ribonuclease A from bovine pancreas

for molecular biology, ≥70 Kunitz units/mg protein, lyophilized

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Synonym(s):
Pancreatic Ribonuclease, RNAsea, RNase A, Ribonucleate 3′-pyrimidinooligonucleotidohydrolase
CAS Number:
9001-99-4
Enzyme Commission number:
3.1.27.5 (BRENDA, IUBMB)
EC Number:
232-646-6
MDL number:
MFCD00132181
UNSPSC Code:
12352204
NACRES:
NA.53

grade

for molecular biology

Quality Level

300

form

lyophilized

specific activity

≥70 Kunitz units/mg protein

mol wt

13.7 kDa
 ~13,700

foreign activity

Exonuclease and endonuclease, none detected
NICKase and DNase, none detected

storage temp.

−20°C

InChI

1S/C9H14N4O3/c10-2-1-8(14)13-7(9(15)16)3-6-4-11-5-12-6/h4-5,7H,1-3,10H2,(H,11,12)(H,13,14)(H,15,16)

InChI key

CQOVPNPJLQNMDC-UHFFFAOYSA-N

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General description

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.
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.

Application

  • 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.
Ribonuclease A from bovine pancreas has been used:
  • for plasmid DNA purification
  • to digest and avoid staining of double-stranded RNA during staining of nuclei
  • in immunoperoxidase and immunofluorescence detection
  • to treat cells prior to flow cytometry
  • as a component of post-hybridization buffer for washing FFPE tissue sections post in-situ hybridization
  • to treat cells during the preparation of outer membrane protein (OMP) extracts
  • RNase protection assays
  • Removal of unspecifically bound RNA
  • Analysis of RNA sequences
  • Hydrolysis of RNA contained in protein samples

Features and Benefits

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

Analysis Note

Protein determined by E.

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.

Application

Product No.
Description
Pricing

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G5663

Glutathione S-Transferase from E. coli, recombinant, expressed in E. coli, buffered aqueous solution

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P6911

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LIG1

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Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Information

动植物源性产品

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. What is the solubility of Product R6513, Ribonuclease A from bovine pancreas (RNase A)?

    Stock solutions of RNase A can be made in 10 mM Tris-HCl, pH 7.5, 15 mM NaCl at a concentration of 10 mg/mL.

  3. What is the shelf life of Product R6513, Ribonuclease A from bovine pancreas (RNase A)? 

    This product is stable for at least 3 years when stored properly at -20 °C.

  4. How are solutions of Product R6513, Ribonuclease A from bovine pancreas (RNase A), stored?

    RNase A solutions can be stored, aliquoted and frozen at -20 °C.

  5. Do I need to boil Product R6513, Ribonuclease A from bovine pancreas (RNase A), to inactivate residual DNases?

    Boiling of stock solutions to inactivate residual DNase is not necessary and may cause precipitation of RNase with loss of enzymatic activity. 

  6. What working concentration of Product R6513, Ribonuclease A from bovine pancreas (RNase A), do you suggest?

    We suggest using RNase A at a concentration of 10 ug/ml.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  9. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  10. Why is RNase A activity not listed as micromoles per minute, and what is a Kunitz unit?

    The Kunitz unit was created to reflect the non-linear nature of the RNAse enzymatic reaction. As the reaction progresses the activity changes due to RNA as a heterogeneous polymeric substrate; therefore, the unit of moles or micomoles per minute becomes inaccurate. As such, our enzymatic assay describes a unit that varies throughout the reaction: One unit will cause a decrease of 100% per minute in the value of E0 - Ef at pH 5.0 at 25°C. Ef is determined in the RNAse control step, while E0 reflects the current reaction rate.

  11. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

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Protocols

Enzymatic Assay of Ribonuclease A (EC 3.1.27.5)

This procedure may be used for determination of Ribonuclease A (RNase A) activity.

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