D5319

Deoxyribonuclease I bovine

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

• Synonim(y): DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• Numer EC enzymu: 3.1.21.1 (BRENDA, IUBMB)
• Numer MDL: MFCD00130918
• Kod UNSPSC: 12352204
• NACRES: NA.54

Właściwości

• pochodzenie biologiczne: bovine
• Poziom jakości: 200
• rekombinowane: expressed in Pichia pastoris
• Próba: ≥95%
• Formularz: buffered aqueous glycerol solution
• aktywność właściwa: ≥5,000 units/mg protein
• masa cząsteczkowa: ~39 kDa
• metody: DNA extraction: suitable
• przydatność: suitable for molecular biology
• Zastosowanie: diagnostic assay manufacturing
• obecność zanieczyszczeń: RNAse and protease, free
• Warunki transportu: dry ice
• temp. przechowywania: −20°C

Opis

Zastosowanie

DNAse I from Sigma was used to treat nuclear lysate to obtain single nucleosomes in a study.[1] The enzyme has also been for the preparation and harvest of mice mammary glands.[2]

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.

Działania biochem./fizjol.

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.[3] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[4] and actin[5] 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.

Cechy i korzyści

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

Definicja jednostki

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

Postać fizyczna

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

Uwaga dotycząca przygotowania

Produced without using any animal cells or animal derived materials.

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Informacja o środkach bezpieczeństwa

• Kod klasy składowania: 10 - Combustible liquids
• Klasa zagrożenia wodnego (WGK): WGK 1
• Temperatura zapłonu (°F): Not applicable
• Temperatura zapłonu (°C): Not applicable