D4527

Deoxyribonuclease I from bovine pancreas

Type II, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein

• Synonim(y): DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• Numer CAS: 9003-98-9
• Numer EC enzymu: 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
• typ: Type II
• Postać: lyophilized powder
• aktywność właściwa: ≥2,000 units/mg protein
• masa cząsteczkowa: ~31 kDa
• oczyszczone przez: chromatography
• skład: Protein, ≥80%
• metody: DNA extraction: suitable
• rozpuszczalność: 0.15 M NaCl: soluble 5.0 mg/mL, clear
• przydatność: suitable for molecular biology
• Zastosowanie: diagnostic assay manufacturing; diagnostic assay manufacturing
• obecność zanieczyszczeń: Chymotrypsin ≤0.01%; Protease ≤0.005%; RNase ≤0.01%
• Warunki transportu: wet ice
• temp. przechowywania: −20°C

Opis

Zastosowanie

DNAse I is used to nick DNA, as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used in the nuclease stock along with RNAse during cell lysate preparation of Madin-Darby canine kidney (MDCK) II cell lines. It has also been used during RNA extraction from Sindbis virus.

Deoxyribonuclease I from bovine pancreas has been used for the identification, localization and expression of two novel human genes similar to deoxyribonuclease I. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, and trypsin from cattle pancreas.

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⁺. 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. 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

Effective for nicking DNA and removal of DNA from protein samples

Definicja jednostki

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

Postać fizyczna

Contains calcium chloride

Uwaga dotycząca przygotowania

The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 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