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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
D4263D4527D5025
impurities

endotoxin, tested

impurities

-

impurities

-

impurities

-

Quality Level

300

Quality Level

200

Quality Level

300

Quality Level

300

form

lyophilized powder

form

powder

form

lyophilized powder

form

lyophilized 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

foreign activity

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

foreign activity

-

foreign activity

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

foreign activity

Chymotrypsin ≤0.5%, Protease ≤0.05%, RNase ≤0.02%

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

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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Stability of DNase I in footprinting experiments.
B Ward et al.
Nucleic acids research, 16(17), 8724-8724 (1988-09-12)
Enzymes of Molecular Biology
Weir, A. F.
Methods in Molecular Biology, 16 (1993)
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
E Lazarides et al.
Proceedings of the National Academy of Sciences of the United States of America, 71(12), 4742-4746 (1974-12-01)
Various tissues and cells in culture contain a specific inhibitor of DNase I (EC 3.1.4.5). In this paper evidence is presented that this inhibitor is actin, one of the major structural proteins of muscle and nonmuscle cells. (a) The inhibitor
T Liao
The Journal of biological chemistry, 250(10), 3831-3836 (1975-05-25)
In the course of experiments on the role of the COOH-terminal residues in pancreatic deoxyribonuclease, we undertook to ascertain whether the presence of sodium dodecyl sulfate would render the normally unavailable terminus susceptible to hydrolysis by carboxypeptidase A. When DNase

Protocols

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

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