MilliporeSigma
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D4527

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

Type II, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein

Synonym(s):
DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

type

Type II

Quality Level

form

lyophilized powder

specific activity

≥2,000 units/mg protein

mol wt

~31 kDa

purified by

chromatography

composition

Protein, ≥80%

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

foreign activity

Chymotrypsin ≤0.01%
Protease ≤0.005%
RNase ≤0.01%

shipped in

wet ice

storage temp.

−20°C

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Application

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 nuclease stock along with RNAse during cell lysate preparation of Madin-Darby canine kidney (MDCK) II cell lines. It has also been used during RNA extraction from Sindbis virus.
Deoxyribonuclease I from bovine pancreas has been used for the identification, localization and expression of two novel human genes similar to deoxyribonuclease I. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, and trypsin from cattle pancreas.
Used for the removal of DNA from protein samples.

Packaging

10000 units in glass bottle
20000, 40000, 200000, 500000 units in poly bottle

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 respectively. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Unit Definition

One Kunitz unit will produce a change in A260 of 0.001 per minute per ml at pH 5.0 at 25 °C using DNA, Type I or III, as the substrate.

Physical form

Contains calcium chloride

Preparation Note

The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 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.

Analysis Note

Protein determined by biuret.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

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Certificate of Origin

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Technical Information

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