Deoxyribonuclease I from bovine pancreas Type II-S, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein Sigma D4513
По запросу
- Производитель: Sigma
- Каталожный номер:
D4513 - CAS номер : 9003-98-9
Analysis Note
Protein determined by biuret.
Application
Used for the removal of DNA from protein samples.
DNAse I from Sigma has been compared with human urine-derived interleukin 1 inhibitor for the ability to hydrolyze [14C]DNA [14C]DNA.1 It has also been used to cleave a 139 base pair Hind III/Nci I restriction fragment to investigate the stability of the enzyme for use in footprinting experiments. DNase I is widely used as a footprinting agent for studying drug and protein binding to DNA.2
Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, and trypsin from cattle pancreas. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the isolation and further characterization of carp liver DNase.
Preparation Note
Derived from New Zealand-sourced pancreas
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.
Unit Definition
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. [Mg2+] = 4.2 mM.
Physical form
Lyophilized powder containing calcium chloride
Biochem/physiol Actions
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+.3 The pH optimum lies between 7 and 8. Divalent cations such as Mn2+, Ca2+, Co2+, and Zn2+ are activators of the enzyme.4 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.5 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)6 and actin7 are known to inhibit the enzyme activity.
Характеристики реактива | |
storage temp. | −20°C |