MilliporeSigma
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D5025

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

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

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

biological source

bovine pancreas

Quality Level

type

Type IV

form

lyophilized powder

specific activity

≥2,000 Kunitz units/mg protein

mol wt

~31 kDa

purified by

chromatography

composition

Protein, ≥80%

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless

application(s)

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

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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 processing of rat brain tissue. This study showed that axonal growth on astrocytes is not inhibited by oligodendrocytes.[81] 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.[82]
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.
Used for the removal of DNA from protein samples.

Packaging

15000 units in glass bottle
150000, 375000, 750000 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+.[34] 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.[40] 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.[54] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[50] and actin[51] 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.

Physical form

Lyophilized powder containing calcium chloride

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.

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

Certificate of Analysis

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Certificate of Origin

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Technical Information

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