DN25

Deoxyribonuclease I from bovine pancreas

lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein

• Synonim(y): Deoxyribonuclease, DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• CAS Number: 9003-98-9
• Numer WE: 3.1.21.1 (BRENDA, IUBMB)
• Numer WE: 232-667-0
• Numer MDL: MFCD00130918
• Kod UNSPSC: 12352204
• NACRES: NA.54

Właściwości

• pochodzenie biologiczne: bovine pancreas
• Poziom jakości: 300
• Formularz: lyophilized powder
• aktywność właściwa: ≥400 Kunitz units/mg protein
• masa cząsteczkowa: ~31 kDa
• skład: Protein, ≥85%
• metody: DNA extraction: suitable
• rozpuszczalność: 0.15 M NaCl: soluble 5.0 mg/mL, hazy
• przydatność: suitable for molecular biology
• Zastosowanie: diagnostic assay manufacturing; diagnostic assay manufacturing
• obecność zanieczyszczeń: RNase ≤0.02%
• Warunki transportu: wet ice
• temp. przechowywania: −20°C

Opis

Opis ogólny

DNase I, or Deoxyribonuclease I, is an endonuclease isolated from bovine pancreas.

• Our DNase I Digests double str and single stranded DNA into oligo and mononucleotides.
• Bovine pancreatic DNase exists as four isozymes, having isoelectric points for A, B, C and D: 5.22, 4.96, 5.06 and 4.78.3. The predominant form is A, with smaller amounts of B and C, and only minor amount of D.
• DNase I structure resembles the structure of to exonuclease III. It includes two central ß sheets. Each β sheet is composed of six β-strands. This complex of β sheets is surrounded by extensive loop and α-helical regions. This enzyme shares structural similarity to exonuclease III.^[1]

Zastosowanie

• Decreases viscosity providing better yields by removing DNA in primary cell isolation:
• Incorporating labelled bases into DNA: DNA nick
• Radioactive labelling
• Bioprocessing applications: DNA removal
• Eliminating genomic DNA from RNA preparations before RT-PCR
• In vitro transcription
• Nick translation
• DNase footprinting
• Actin regulation of actin polymerization in cells, and cell apoptosis
• UV crosslinking of proteins to nucleic acids
• DNase play a role in the regulation of actin polymerization in cells and is involved in apoptosis process ^[1]

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. In the presence of Mg²⁺, DNAse I cleaves each strand of DNA independently and the cleavage sites are random. Both DNA strands are cleaved at approximately the same site in the presence of Mn²⁺. 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. The pH optimum is found to be between 7 and 8. DNAse I from bovine pancreas consists of four chromatographically distinguishable components, A, B, C, and D, with their molar ratios being 4:1:1 respectively. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Cechy i korzyści

Our Deoxyribonuclease DNAse I,is essentially RNAse free product, which support the product application.

Definicja jednostki

One Kunitz unit will produce a change in A₂₆₀ of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III, as substrate.

Postać fizyczna

Crude preparation, contains calcium chloride

Uwaga dotycząca przygotowania

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 up to five hours at 60 °C between pH 5 and 7, 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.

Komentarz do analizy

Protein determined by biuret.

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Informacja o środkach bezpieczeństwa

• Piktogramy: GHS08
• Hasło ostrzegawcze: Danger
• Zwroty wskazujące rodzaj zagrożenia: H334
• Zwroty wskazujące środki ostrożności: P261 - P284 - P501
• Klasyfikacja zagrożeń: Resp. Sens. 1
• Kod klasy składowania: 11 - Combustible Solids
• Klasa zagrożenia wodnego (WGK): WGK 3
• Temperatura zapłonu (°F): Not applicable
• Temperatura zapłonu (°C): Not applicable
• Środki ochrony indywidualnej: Eyeshields, Gloves, type N95 (US)