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DN25

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein

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

form

lyophilized powder

Quality Level

specific activity

≥400 Kunitz units/mg protein

mol wt

~31 kDa

composition

Protein, ≥85%

solubility

0.15 M NaCl: soluble 5.0 mg/mL, hazy

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

foreign activity

RNase ≤0.02%

shipped in

wet ice

storage temp.

−20°C

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General description

DNase I, or Deoxyribonuclease I, is an endonuclease isolated from bovine pancreas.
  • Our DNase I Digests double str and single stranded DNA into oligo and mononucleotides.
  • Bovine pancreatic DNase exists as four isozymes, having isoelectric points for A, B, C and D: 5.22, 4.96, 5.06 and 4.78.3. The predominant form is A, with smaller amounts of B and C, and only minor amount of D.
  • DNase I structure resembles the structure of to exonuclease III. It includes two central ß sheets. Each β sheet is composed of six β-strands. This complex of β sheets is surrounded by extensive loop and α-helical regions. This enzyme shares structural similarity to exonuclease III.[1]

Application

  • Decreases viscosity providing better yields by removing DNA in primary cell isolation:
  • Incorporating labelled bases into DNA: DNA nick
  • Radioactive labelling
  • Bioprocessing applications: DNA removal
  • Eliminating genomic DNA from RNA preparations before RT-PCR
  • In vitro transcription
  • Nick translation
  • DNase footprinting
  • Actin regulation of actin polymerization in cells, and cell apoptosis
  • UV crosslinking of proteins to nucleic acids
  • DNase play a role in the regulation of actin polymerization in cells and is involved in apoptosis process [1]
Used for the removal of DNA from protein samples.

Packaging

10, 100 mg in poly bottle
1, 5 g 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+. 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. The pH optimum is found to be between 7 and 8. 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.

Features and Benefits

Our Deoxyribonuclease DNAse I,is essentially RNAse free product, which support the product application.

Unit Definition

One Kunitz unit will produce a change in A260 of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III, as substrate.

Physical form

Crude preparation, contains 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

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificate of Analysis

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Certificate of Origin

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Product Information Sheet

More Documents

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