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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

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

bovine pancreas

Quality Level

form

lyophilized powder

specific activity

≥400 Kunitz units/mg protein

mol wt

~31 kDa

composition

Protein, ≥85%

technique(s)

DNA extraction: suitable

solubility

0.15 M NaCl: soluble 5.0 mg/mL, hazy

suitability

suitable for molecular biology

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.

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

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

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Purification of rat spermatogenic cells and preliminary biochemical analysis of these cells.
M L Meistrich et al.
Biology of reproduction, 25(5), 1065-1077 (1981-12-01)
Enzymes of Molecular Biology
Weir, A.F.
Methods in Molecular Biology, 16(2), 2-16 null
Murali Muniraju et al.
Journal of virology, 93(16) (2019-05-31)
Kaposi sarcoma-associated herpesvirus (KSHV) is an emerging pathogen and is the causative infectious agent of Kaposi sarcoma and two malignancies of B cell origin. To date, there is no licensed KSHV vaccine. Development of an effective vaccine against KSHV continues
Comparison of the multiple forms of bovine pancreatic deoxyribonuclease.
J Salnikow et al.
The Journal of biological chemistry, 245(21), 5685-5690 (1970-11-10)
G A Scarborough
Proceedings of the National Academy of Sciences of the United States of America, 73(5), 1485-1488 (1976-05-01)
Biochemicalical evidence is presented which demonstrates that the Neurospora crassa plasma membrane ATPase (ATP phosphohydrolase, EC 3.6.1.3) is an electrogenic pump. The electrical potential across the Neurospora plasma membrane, as monitored by [14C]SCN- uptake by isolated Neurospora plasma membrane vesicles

Protocols

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

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