D5319

Deoxyribonuclease I bovine

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

• Synonyme(s) : DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• Numéro de classification (Commission des enzymes): 3.1.21.1 (BRENDA, IUBMB)
• Numéro MDL: MFCD00130918
• Code UNSPSC : 12352204
• Nomenclature NACRES : NA.54

Propriétés

• Source biologique: bovine
• Niveau de qualité: 200
• Produit recombinant: expressed in Pichia pastoris
• Pureté: ≥95%
• Forme: buffered aqueous glycerol solution
• Activité spécifique: ≥5,000 units/mg protein
• Poids mol.: ~39 kDa
• Technique(s): DNA extraction: suitable
• Adéquation: suitable for molecular biology
• Application(s): diagnostic assay manufacturing
• Activité étrangère: RNAse and protease, free
• Conditions d'expédition: dry ice
• Température de stockage: −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.

Actions biochimiques/physiologiques

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.

Caractéristiques et avantages

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

Définition de l'unité

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

Forme physique

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

Notes préparatoires

Produced without using any animal cells or animal derived materials.

Informations sur la sécurité

• Code de la classe de stockage: 10 - Combustible liquids
• Classe de danger pour l'eau (WGK): WGK 1
• Point d'éclair (°F): Not applicable
• Point d'éclair (°C): Not applicable