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D5025 Sigma

Deoxyribonuclease I from bovine pancreas

Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein

Synonym: DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase

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Properties

Related Categories 3.1.x.x Acting on esters, 3.x.x.x Hydrolases, Apoptosis and Cell Cycle, Application Index, Biochemicals and Reagents,
Quality Level   PREMIUM
type   Type IV
form   lyophilized powder
mol wt   mol wt ~31 kDa
purified by   chromatography
composition   Protein, ≥80%
solubility   0.15 M NaCl: soluble5.0 mg/mL, clear, colorless
Featured Industry   Diagnostic Assay Manufacturing
Diagnostic Assay Manufacturing
foreign activity   Chymotrypsin ≤0.5%
  Protease ≤0.05%
  RNase ≤0.02%
shipped in   wet ice
storage temp.   −20°C

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Analysis Note

Protein determined by biuret.

Application

Used for the removal of DNA from protein samples.

DNAse I is used to nick DNA as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used in the processing of rat brain tissue. This study showed that axonal growth on astrocytes is not inhibited by oligodendrocytes. In another study, thawed fixed samples of E. coli were digested with DNAse I from Sigma along with other enzymes. The digestion was done before permeabilization and staining of the nucleic acids.

Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a two-dimensional zymogram analysis of nucleases in Bacillus subtilis. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effects of minor and major groove-binding drugs and intercalators on the DNA association of minor groove-binding proteins RecA and deoxyribonuclease I.

Packaging

15000 units in glass bottle

150000, 375000, 750000 units in poly bottle

Preparation Note

10 mg/mL solution of DNAse I in 0.15 M NaCl may lose <10% of its activity when stored for a week in aliquots at −20 °C. The same solutions stored in aliquots at 2-8 °C can lose approximately 20% activity. It remains active for up to five hours at 60 °C between pH 5 and 7, and loses activity in <10 minutes at 68 °C. It loses activity at the rate of 6%/hour in acetate buffer (pH 5.0) and tris buffer ((pH 7.2) at 1 mg/mL concentration.

Unit Definition

One Kunitz unit will produce a ΔA260 of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III as substrate.

Physical form

Lyophilized powder containing calcium chloride

Biochem/physiol Actions

DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5′-phosphates. In the presence of Mg2+, DNAse I cleaves each strand of DNA independently and the cleavage sites are random. Both DNA strands are cleaved at approximately the same site in the presence of Mn2+. The pH optimum is found to be between 7 and 8. Divalent cations such as Mn2+, Ca2+, Co2+, and Zn2+ are activators of the enzyme. A concentration of 5 mM Ca2+ stabilizes the enzyme against proteolytic digestion. DNAse I from bovine pancreas consists of four chromatographically distinguishable components, A, B, C, and D, with their molar ratios being 4:1:1. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Price and Availability


All labs need water

Biomedical Applications
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
RTECS 
RF0750000

Frequently Asked Questions

How should I prepare a solution of Product D5025, DNAse I?
This enzyme can be reconstituted in 0.15 M NaCl at a concentration of 10 mg/ml. To view this information in the D5025 data sheet, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/3/d5025dat.Par.0001.File.tmp/d5025dat.pdf
Are stock solutions of Product D5025, Deoxyribonuclease I (DNase I), stable?
Solutions of DNAse I (10 mg/ml) in 0.15 M NaCl may lose 10% of its activity when stored for a week in aliquots at -20°C. The same solutions stored in aliquots at 2 - 8 deg. C. lose approximately 20% activity. To view this information on the D5025 data sheet click on the following link: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/3/d5025dat.Par.0001.File.tmp/d5025dat.pdfDNAse
How do Deoxyribonuclease I (DNase I), products D5025 and D4527  differ from each other?
Both of these products have the same specific activity (>2000 units/mg protein) for Dnase I. The enzyme impurity levels for each, however, are different. D5025 has the following impurity specification: Chymotrypsin (<0.5%), Protease (<0.05%), and RNase (<0.02%). The impurity specifications for D4527 are: Chymotrypsin (<0.01%), Protease (<0.005%), and RNase (<0.01%). To view this information click on the following links: http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=D5025|SIGMA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC    and http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=D4527|SIGMA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC
How can I convert units of Product D5025, Deoxyribonuclease I (DNase I), to mass?
The mg of solid for each unit package size will be indicated on the label of the package. For a particular lot, you can calculate the specific activity in units per mg solid by multiplying the specific activity in units per mg protein by the percent protein divided by 100. To view this lot specific information, click on the following link:  http://www.sigmaaldrich.com/catalog/search/ProductDetail/SIGMA/D5025. Under related information, enter your lot number in the certificate of analysis field.
What concentration of Product D5025, Deoxyribonuclease I (DNase I),  is required to remove DNA from solutions?
DNA can be removed from preparations by incubation with 20 - 50 ug DNAse I in the presence of 50 mM Tris-HCl buffer, pH 7.5, 10 mM magnesium chloride, at 37 deg. C. for 60 min.
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.
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.
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Protocols & Articles

Protocols

Enzymatic Assay of Deoxyribonuclease I (EC 3.1.21.1)

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I at Sigma-Aldrich St. Louis.

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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