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D4513

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

Type II-S, 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

sterility

sterile-filtered

type

Type II-S

form

lyophilized powder

specific activity

≥2,000 units/mg protein

mol wt

~31 kDa

purified by

chromatography

composition

Protein, ≥80%

packaging

vial of ≥10.0 mg protein

impurities

endotoxin, tested

solubility

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

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

foreign activity

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

storage temp.

−20°C

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1 of 4

This Item
D4263D5025DN25
form

lyophilized powder

form

powder

form

lyophilized powder

form

lyophilized powder

specific activity

≥2,000 units/mg protein

specific activity

-

specific activity

≥2,000 Kunitz units/mg protein

specific activity

≥400 Kunitz units/mg protein

mol wt

~31 kDa

mol wt

~31 kDa

mol wt

~31 kDa

mol wt

~31 kDa

purified by

chromatography

purified by

chromatography

purified by

chromatography

purified by

-

impurities

endotoxin, tested

impurities

-

impurities

-

impurities

-

Application

DNAse I from Sigma has been compared with human urine-derived interleukin 1 inhibitor for the ability to hydrolyze [14C]DNA [14C]DNA. It has also been used to cleave a 139 base pair Hind III/Nci I restriction fragment to investigate the stability of the enzyme for use in footprinting experiments. DNase I is widely used as a footprinting agent for studying drug and protein binding to DNA.
Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, and trypsin from cattle pancreas. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the isolation and further characterization of carp liver DNase.
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 lies 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 ΔA260 of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III as substrate. (1 unit = 1 Kunitz unit)

Physical form

Lyophilized powder containing 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

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

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Enzymes of Molecular Biology
Weir, A. F.
Methods in Molecular Biology, 16 (1993)
Stability of DNase I in footprinting experiments.
B Ward et al.
Nucleic acids research, 16(17), 8724-8724 (1988-09-12)
Sambrook, J., and Russell, D.W.
Molecular Cloning: A Laboratory Manual, 2(2), 5-5 (2001)
D L Rosenstreich et al.
The Journal of experimental medicine, 168(5), 1767-1779 (1988-11-01)
We have previously reported that the urine of febrile humans contained large quantities of an inhibitor of IL-1-induced murine thymocyte proliferation that was a glycoprotein between 30 and 40 kD in size. In the present study this factor has been
Daniel Engelbertsen et al.
Scientific reports, 9(1), 10608-10608 (2019-07-25)
CD47, also known as integrin-associated protein (IAP), is a transmembrane protein with multiple biological functions including regulation of efferocytosis and leukocyte trafficking. In this study we investigated the effect of CD47-deficiency on atherosclerosis using a model of adeno-associated virus (AAV)-induced

Protocols

Enzymatic Assay of Deoxyribonuclease I (EC 3.1.21.1)

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

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