D5025

Deoxyribonuclease I from bovine pancreas

Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein

• Szinonimák: DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• CAS-szám: 9003-98-9
• Enzyme Commission szám: 3.1.21.1 (BRENDA, IUBMB)
• EC-szám: 232-667-0
• MDL-szám: MFCD00130918
• UNSPSC kód: 12352204
• NACRES: NA.54

Tulajdonságok

• biológiai forrás: bovine pancreas
• Minőségi szint: 300
• típus: Type IV
• form: lyophilized powder
• specifikus aktivitás: ≥2,000 Kunitz units/mg protein
• molekulatömeg: ~31 kDa
• tisztítva: chromatography
• összetétel: Protein, ≥80%
• technika/technikák: DNA purification: suitable
• oldhatóság: 0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless
• alkalmasság: suitable for molecular biology
• alkalmazás(ok): diagnostic assay manufacturing; diagnostic assay manufacturing
• idegen aktivitás: Chymotrypsin ≤0.5%; Protease ≤0.05%; RNase ≤0.02%
• kiszállítva: wet ice
• tárolási hőmérséklet: −20°C

Leírás

Alkalmazás

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 processing of rat brain tissue. This study showed that axonal growth on astrocytes is not inhibited by oligodendrocytes. In another study, thawed fixed samples of E. coli were digested with DNAse I from Sigma along with other enzymes. The digestion was done before permeabilization and staining of the nucleic acids.

Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a two-dimensional zymogram analysis of nucleases in Bacillus subtilis. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effects of minor and major groove-binding drugs and intercalators on the DNA association of minor groove-binding proteins RecA and deoxyribonuclease I.

Used for the removal of DNA from protein samples.

Biokémiai/fiziológiai hatások

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. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Egység definíció

One Kunitz unit will produce a ΔA₂₆₀ of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III as substrate.

Fizikai forma

Lyophilized powder containing calcium chloride

Elkészítési megjegyzés

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.

Analízis megjegyzés

Protein determined by biuret.

Biztonsági adatok

• Piktogramok: GHS08
• Figyelmeztetés: Danger
• Figyelmeztető mondatok: H334
• Óvintézkedésre vonatkozó mondatok: P261 - P284 - P501
• Veszélyességi osztályok: Resp. Sens. 1
• Tárolási osztály kódja: 11 - Combustible Solids
• WGK: WGK 3
• Lobbanási pont (F): Not applicable
• Lobbanási pont (C): Not applicable