This product is soluble in 0.15 M NaCl at 5 mg/mL. Solutions prepared at 10 mg/mL in 0.15 M NaCl may lose up to 10% activity when stored for 1 week in aliquots at –20 °C. The same solutions stored in aliquots at 2-8 °C can lose up to 20% activity. Please see the link below to review the product datasheet for additional information:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/233/818/dn25pis-ms.pdf
DN25
Deoxyribonuclease I from bovine pancreas
Deoxyribonuclease I from bovine pancreas
1 of 3 reviewers received a sample product or took part in a promotion
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
Pricing and availability is not currently available.
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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+.[1] 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.[2] 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.[3] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[4] and actin[5] 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.
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
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
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HOW CAN I PRESERVE DN25 DILUTED and filtered in culture medium? 4C or -20 C?. It will lose activity?
1 answer-
Helpful?
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What is the recommended method for long-term storage of the 1 gram bottle of DN25 in solution?
1 answer-
The user bulletin provides guidance on the storage of DNase I once reconstituted. When stored in aliquots at -20°C, solutions of DNase I at 10 mg/mL in 0.15 M NaCl may lose 10% of activity over a week, while storing the same solutions at 2–8°C can result in a 20% activity loss. Alternatively, maintaining solutions at a minimum concentration of 1 mg protein/mL with 5 mM calcium chloride at -20°C can retain 90% activity for at least a year. Solutions with 0.1 mg protein/mL are less stable and may need gelatin as a stabilizer.
For applications not affected by glycerol, two storage buffer options are available as they do not freeze at -20°C:
• 20 mM sodium acetate (pH 6.5) with 5 mM CaCl2, 0.1 mM phenylmethylsulfonyl fluoride (PMSF), and 50% (v/v) glycerol, containing DNase I at 5 mg/mL.
• 10 mM Tris-HCl (pH 7.5) with 50 mM NaCl, 10 mM MgCl2, 1 mM DTT, and 50% (v/v) glycerol, with DNase I at 2 mg/mL.Helpful?
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Hi I use the DNase during isolation of cells from lung tissue. Do I need to filter the DNase solution before usage?
1 answer-
This is not a sterile product. If sterility is required due to the specific application, solutions may be filter sterilized before use. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/233/818/dn25pis-ms.pdfHelpful?
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I use the DNase when I isolate lung cells from lung. Do I need to filter them before I use? I will culture the isolated lung cells.
1 answer-
Yes, please filter the DNase solution before use in cell culture, preferably with a 0.22 um filter. The powder is not sterilized as provided.
Helpful?
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