D5319

Deoxyribonuclease I bovine

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

• Sinónimos: DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• Comisión internacional de enzimas: 3.1.21.1 (BRENDA, IUBMB)
• Número MDL: MFCD00130918
• Código UNSPSC: 12352204
• NACRES: NA.54

Propiedades

• origen biológico: bovine
• Nivel de calidad: 200
• recombinante: expressed in Pichia pastoris
• Ensayo: ≥95%
• Formulario: buffered aqueous glycerol solution
• actividad específica: ≥5,000 units/mg protein
• mol peso: ~39 kDa
• técnicas: DNA extraction: suitable
• idoneidad: suitable for molecular biology
• aplicaciones: diagnostic assay manufacturing
• actividad extraña: RNAse and protease, free
• Condiciones de envío: dry ice
• temp. de almacenamiento: −20°C

Descripción

Aplicación

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.

Acciones bioquímicas o fisiológicas

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.

Características y beneficios

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

Definición de unidad

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

Forma física

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

Nota de preparación

Produced without using any animal cells or animal derived materials.

Información de seguridad

• Código de clase de almacenamiento: 10 - Combustible liquids
• Clase de riesgo para el agua (WGK): WGK 1
• Punto de inflamabilidad (°F): Not applicable
• Punto de inflamabilidad (°C): Not applicable