As cited in the product listing and datasheet, this product in solution shows an approximate loss of activity of 10% when stored in aliquots at −20 °C for 1 week. Studies at lower dilutions have not been performed. Typically, solutions stored at higher concentrations are the most stable.
D5025
Deoxyribonuclease I from bovine pancreas
Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
Synonym(s):
DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
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About This Item
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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%
technique(s)
DNA purification: suitable
solubility
0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless
suitability
suitable for molecular biology
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.[1] 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.[2]
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.
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+.[3] 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.[4] 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.[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[6] and actin[7] 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.
inhibitor
Product No.
Description
Pricing
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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Protocols
To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.
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Good afternoon, if the stock concentration of 10mg/ml may loose 10% of its activity after one week, I was wondering if it is okey regarding stability to keep 500ug/ml aliquots at -20ºC for several weeks.
1 answer-
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Is Kunitz unit same as unit here? I have seen some papers and they have U/ml for DNase. How can i conver it?
1 answer-
Product D5025, Deoxyribonuclease I is given in Kunitz units. This product is a powder. When reconstituting the product with a solution, for example 15,000 Kunitz units in 1 ml of 0.15 M NaCl the end-user will have 15,000 Kunitz units/ml.
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What concentration of Product D5025, Deoxyribonuclease I (DNase I), is required to remove DNA from solutions?
1 answer-
DNA can be removed from preparations by incubation with 20 - 50 ug DNAse I in the presence of 50 mM Tris-HCl buffer, pH 7.5, 10 mM magnesium chloride, at 37°C. for 60 min.
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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How should I prepare a solution of Product D5025, DNAse I?
1 answer-
This enzyme can be reconstituted in 0.15 M NaCl at a concentration of 10 mg/ml.
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How do Deoxyribonuclease I (DNase I), products D5025 and D4527 differ from each other?
1 answer-
Both of these products have the same specific activity (>2000 units/mg protein) for Dnase I. The enzyme impurity levels for each, however, are different. D5025 has the following impurity specification: Chymotrypsin (<0.5%), Protease (<0.05%), and RNase (<0.02%). The impurity specifications for D4527 are: Chymotrypsin (<0.01%), Protease (<0.005%), and RNase (<0.01%).
Helpful?
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Are stock solutions of Product D5025, Deoxyribonuclease I (DNase I), stable?
1 answer-
Solutions of DNAse I (10 mg/ml) 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. lose approximately 20% activity.
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How can I convert units of Product D5025, Deoxyribonuclease I (DNase I), to mass?
1 answer-
The mg of solid for each unit package size will be indicated on the label of the package. For a particular lot, you can calculate the specific activity in units per mg solid by multiplying the specific activity in units per mg protein by the percent protein divided by 100.
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