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D4527

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

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

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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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1 of 4

This Item
D4513DN25D4263
form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

powder

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear(lit.)

solubility

0.15 M NaCl: soluble 5.0 mg/mL, hazy

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

foreign activity

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

foreign activity

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

foreign activity

RNase ≤0.02%

foreign activity

-

shipped in

wet ice

shipped in

-

shipped in

wet ice

shipped in

-

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.

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.

Features and Benefits

Effective for nicking DNA and removal of DNA from protein samples

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

Precautionary 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


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Enzymes of Molecular Biology
Weir, A. F.
Methods in Molecular Biology, 16 (1993)
Y Shirako et al.
Journal of virology, 68(3), 1874-1885 (1994-03-01)
Nonstructural proteins of Sindbis virus, nsP1, nsP2, nsP3, and nsP4, as well as intermediate polyproteins, are produced from two precursor polyproteins, P123 and P1234, by a proteolytic enzyme encoded in the C-terminal half of nsP2. We studied the requirements for
David B N Lee et al.
American journal of physiology. Renal physiology, 287(3), F481-F491 (2004-04-29)
The tight junction has been characterized as a domain of focal fusions of the exoplasmic leaflets of the lipid bilayers from adjacent epithelial cells. Approximating membranes to within fusion distance is a thermodynamically unfavorable process and requires the participation of
Tianxu Han et al.
EMBO reports, 17(12), 1814-1828 (2016-11-01)
Hematopoietic stem cells (HSCs) are capable of giving rise to all blood cell lineages throughout adulthood, and the generation of engraftable HSCs from human pluripotent stem cells is a major goal for regenerative medicine. Here, we describe a functional genome-wide
Ben L Callif et al.
Molecular and cellular neurosciences, 80, 170-179 (2017-01-23)
Axon growth is coordinated by multiple interacting proteins that remain incompletely characterized. High content screening (HCS), in which manipulation of candidate genes is combined with rapid image analysis of phenotypic effects, has emerged as a powerful technique to identify key

Protocols

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

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