D5319

Deoxyribonuclease I bovine

recombinant, expressed in Pichia pastoris, buffered aqueous glycerol solution, ≥5,000 units/mg protein

• Synonym(s): DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• UNSPSC Code: 12352204
• NACRES: NA.54
• EC Number: 3.1.21.1 (BRENDA, IUBMB)
• MDL number: MFCD00130918
• Specific activity: ≥5,000 units/mg protein
• Assay: ≥95%
• Biological source: bovine
• Recombinant: expressed in Pichia pastoris

Properties

• biological source: bovine
• Quality Segment: 200
• recombinant: expressed in Pichia pastoris
• assay: ≥95%
• form: buffered aqueous glycerol solution
• specific activity: ≥5,000 units/mg protein
• mol wt: ~39 kDa
• technique(s): DNA extraction: suitable
• input: general use sample(s)
• suitability: suitable for molecular biology
• application(s): diagnostic assay manufacturing
• foreign activity: RNAse and protease, free
• shipped in: dry ice
• storage temp.: −20°C

Description

Application

DNAse I from Sigma was used to treat nuclear lysate to obtain single nucleosomes in a study. The enzyme has also been for the preparation and harvest of mice mammary glands.

Deoxyribonuclease I bovine has been used in a study to compare the initial actions of spleen deoxyribonuclease and pancreatic deoxyribonuclease. Deoxyribonuclease I bovine has also been used in a study to investigate deoxythymidine 3′, 5′-di-p-nitrophenyl phosphate as a synthetic substrate for bovine pancreatic deoxyribonuclease.

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

Physical form

Supplied as a solution in 4 mg/ml glycine pH 5.0, 5 mM calcium acetate and 50% glycerol

Preparation Note

Produced without using any animal cells or animal derived materials.

Other Notes

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

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable