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D4263 Sigma

Deoxyribonuclease I from bovine pancreas

Standardized vial containing 2,000 Kunitz units of DNase I (D4527), vial of ≥0.25 mg total 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,
form   powder
mol wt   mol wt ~31 kDa
purified by   chromatography
packaging   vial of ≥0.25 mg total protein
solubility   0.15 M NaCl: soluble5.0 mg/mL, clear
Featured Industry   Diagnostic Assay Manufacturing
Diagnostic Assay Manufacturing
storage temp.   −20°C

Description

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. DNAse I from Sigma has been used along with other enzymes for tumor harvest and dissociation, during the isolation and molecular characterization of cancer stem cells in MMTV-Wnt-1 murine breast tumors.

Deoxyribonuclease I from bovine pancreas has been used in a study to determine that mammalian deoxyribonucleases I are classified into three types based on differences in their tissue concentrations. Deoxyribonuclease I from bovine pancreas has also been used in a study to compare the three primary structures of deoxyribonuclease isolated from bovine, ovine, and porcine pancreas.

Packaging

1 vial in serum bottle

5 vials in serum bottle

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.

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.

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 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.

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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