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D2821

Sigma-Aldrich

Deoxyribonuclease I bovine

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

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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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This Item
D5319A6237D5307
Apyrase from potato recombinant, expressed in Pichia pastoris, ATPase ≥1000 units/mg protein, lyophilized powder

Sigma-Aldrich

A6237

Apyrase from potato

form

lyophilized powder

form

buffered aqueous glycerol solution

form

lyophilized powder

form

liquid

specific activity

≥4,000 units/mg protein

specific activity

≥5,000 units/mg protein

specific activity

-

specific activity

-

mol wt

~39 kDa

mol wt

~39 kDa

mol wt

-

mol wt

-

solubility

H2O: soluble (pH 4.0-9.0)

solubility

-

solubility

-

solubility

-

foreign activity

RNAse and protease, free

foreign activity

RNAse and protease, free

foreign activity

-

foreign activity

-

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, complete RNA lysis buffer, cell lysis buffer for testing the expression of recombinant tagged protein.
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.
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. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin 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

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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