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R6513 Sigma

Ribonuclease A from bovine pancreas

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

Synonym: Pancreatic Ribonuclease, RNase A, Ribonuclease I, Ribonucleate 3′-pyrimidinooligonucleotidohydrolase

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Description

Analysis Note

Protein determined by E1%/280.

Features and Benefits

• RNase protection assays
• Remove unspecifically bound RNA
• Analysis of RNA sequences
• Hydrolyze RNA contained in protein samples
• Purification of DNA

General description

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.

Application

Ribonuclease A is used to remove RNA from DNA plasmid preparations and protein samples. It is also used in RNA sequence analysis and protection assays.

Price and Availability


Biomedical Applications
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
RTECS 
RF0760000

Frequently Asked Questions

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. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
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. The solubility information can be viewed on the data sheet; here is the link to that document:   http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/5/r6513dat.Par.0001.File.tmp/r6513dat.pdf
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. The shelf life information can be viewed on the Specification Sheet by accessing the following link:  http://www.sigmaaldrich.com/catalog/DataSheetPage.do?brandKey=SIGMA&symbol=R6513
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.
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.  The solubility information can be viewed on lot specific Datasheet by accessing the following link: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/5/r6513dat.Par.0001.File.tmp/r6513dat.pdf
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.  The working concentration information can be viewed on the Product Datasheet by accessing the following link:   http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/5/r6513dat.Par.0001.File.tmp/r6513dat.pdf
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
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. 
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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.
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Protocols & Articles

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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. The Spectrophotometric Stop Rate Determination [Absorbance at 300 nm (A300), Light path = 1 cm] is based on the foll...

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Peer-Reviewed Papers
15

References

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