MilliporeSigma
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D2821

Sigma-Aldrich

Deoxyribonuclease I bovine

recombinant, expressed in Pichia pastoris, lyophilized powder, RNAse and protease, free

Synonym(s):
DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
Enzyme Commission number:
MDL number:
NACRES:
NA.54

recombinant

expressed in Pichia pastoris

Quality Level

form

lyophilized powder

specific activity

≥4,000 units/mg protein

mol wt

~39 kDa

solubility

H2O: soluble (pH 4.0-9.0)

foreign activity

RNAse and protease, free

storage temp.

2-8°C

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Application

Deoxyribonuclease I bovine has been used in a study to investigate the inhibition of DNA polymerase by extracts of rat liver. Deoxyribonuclease I bovine has also been used in a study to investigate the effects of ionic strength on enzymic activity.
Deoxyribonuclease I bovine has been used in the preparation of cold cell lysis buffer,[1] complete RNA lysis buffer,[2] cell lysis buffer for testing the expression of recombinant tagged protein.[3]
The enzyme from Sigma has been used for the digestion of DNA extracted from Agave spp. clones. The enzyme was used during a study that investigated the effect of epigenetic changes on the regulatory expression of KNOTTED1-like HOMEOBOX (KNOX) transcription factors.[4]
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. 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.[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[6] and actin[7] are known to inhibit the enzyme activity.
Digests single- and double-stranded DNA to a mixture of mono- and oligonucleotides carrying 5′ phosphates and 3′ OH termini. This catalytic activity is divalent ion-dependent. In the presence of Mg2+, DNase I hydrolyzes each strand of double-stranded DNA randomly and independently. In the presence of Mn2+, both strands can be cleaved.

Features and Benefits

  • RNA purification by removing DNA
  • Prepare DNA for nick translation1
  • Footprinting assays to determine DNA-protein interactions2

Unit Definition

One unit will produce a ΔA260 of 0.001 per min per mL reaction mixture using calf thymus DNA at pH 5.0 and 25°C

Physical form

supplied as a lyophilized powder containing glycine as a stabilizer

Preparation Note

Produced without using any animal cells or animal derived materials.
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 upto five hours at 60 °C 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.

Storage Class Code

13 - Non Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

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Certificate of Origin

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