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D5025 Sigma

Deoxyribonuclease I from bovine pancreas

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

Synonym: DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase

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Properties

Related Categories 3.1.x.x Acting on esters, 3.x.x.x Hydrolases, Apoptosis and Cell Cycle, Application Index, Biochemicals and Reagents,
Quality Level   PREMIUM
type   Type IV
form   lyophilized powder
mol wt   mol wt ~31 kDa
purified by   chromatography
composition   Protein, ≥80%
solubility   0.15 M NaCl: soluble5.0 mg/mL, clear, colorless
Featured Industry   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

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Analysis Note

Protein determined by biuret.

Application

Used for the removal of DNA from protein samples.

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. 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.

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.

Packaging

15000 units in glass bottle

150000, 375000, 750000 units in poly bottle

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.

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

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. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Price and Availability


Flow Cytometry Instruments

All labs need water
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
RTECS 
RF0750000
Protocols & Articles

Protocols

Enzymatic Assay of Deoxyribonuclease I (EC 3.1.21.1)

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I at Sigma-Aldrich St. Louis.

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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